Inhibitors of human immunodeficiency virus replication

ABSTRACT

Compounds of formula I: 
     
       
         
         
             
             
         
       
     
     wherein R 4 , R 6  and R 7  are defined herein, are useful as inhibitors of HIV replication.

RELATED APPLICATIONS

This application claims benefit of U.S. Ser. No. 60/988,686, filed Nov.16, 2007, which is herein incorporated by reference.

FIELD OF THE INVENTION

The present invention relates to compounds, compositions and methods forthe treatment of human immunodeficiency virus (HIV) infection. Inparticular, the present invention provides novel inhibitors of HIVreplication, pharmaceutical compositions containing such compounds andmethods for using these compounds in the treatment of HIV infection.More specifically, the present invention provides novel inhibitors ofthe HIV integrase enzyme, pharmaceutical compositions containing suchcompounds and methods for using these compounds to reduce HIVreplication and in the treatment of HIV infection.

BACKGROUND OF THE INVENTION

Acquired immune deficiency syndrome (AIDS) is caused by the humanimmunodeficiency virus (HIV), particularly the HIV-1 strain. Mostcurrently approved therapies for HIV infection target the viral reversetranscriptase and protease enzymes. There is additionally one approveddrug targeting gp41 to inhibit viral entry and one approved drugtargeting the integrase enzyme. Within the reverse transcriptaseinhibitor and protease inhibitor classes, resistance of HIV to existingdrugs is a problem. Therefore, it is important to discover and developnew antiretroviral compounds.

The inherent genetic variation within HIV has led to the identificationof many HIV mutants, commonly referred to as variants, which exhibitaltered drug susceptibility. On the integrase enzyme, residues 124 and125 are recognized as highly variable across the HIV-1 virus frominfected patients found in major market and developing countries. Theapproximate prevalence of these integrase variants are Thr124/Thr125(44%), Ala124/Thr125 (17%), Ala124/Ala125 (16%), Thr124/Ala125 (10%),Asn124/Thr125 (6%), and Asn124/Ala125 (1%) for viruses sequenced frommajor market countries reported in the Los Alamos database(http://www.hiv.lanl.gov/content/hiv-db). These integrase variants maybe generated using known methods in the art and publicly availablepolypeptide sequences for the integrase enzyme, for example from theNL4.3 strain of HIV-1 integrase (SEQ ID NO: 1).

SUMMARY OF THE INVENTION

The present invention provides a novel series of compounds havinginhibitory activity against HIV replication. The compounds of thepresent invention have an affinity for the HIV integrase enzyme.Therefore, the compounds of the invention may be used to inhibit theactivity of HIV integrase and may be used to reduce HIV replication. Thecompounds of the invention exhibit at least one of the followingsurprising advantages:

-   -   unexpectedly good activity in a cell-based HIV-1 replication        assay in four of the major integrase variants at residues        124/125 (Thr124/Thr125, Ala124/Thr125, Ala124/Ala125 and        Thr124/Ala125); and/or    -   unexpectedly good activity in a cell-based HIV-1 replication        assay in all six of the major integrase variants at residues        124/125 (Thr124/Thr125, Ala124/Thr125, Ala124/Ala125,        Thr124/Ala125, Asn124/Thr125, and Asn124/Ala125); and/or    -   unexpectedly good pharmacological properties.

The compounds of the invention exhibit unexpectedly good potency againstfour of major integrase variants at the 124/125 residues (˜>85% naturalabundance) and/or all six of the major integrase variants at the 124/125residues. The implication of this unexpectedly good potency observedacross the aforementioned 124/125 variable residues is that some HIVinfected patients, who carry a virus with 124/125 variant residues ofintegrase and who have a pre-existing anti-viral resistance againstdrugs of this class, may be expected to respond to the compounds of theinvention.

Further objects of this invention arise for the one skilled in the artfrom the following description and the examples.

One aspect of the invention provides an isomer, racemate, enantiomer ordiasteriomer of compounds of formula (I):

wherein

-   R⁴ is aryl or Het, wherein each of the aryl and Het is optionally    substituted with 1 to 3 substituents each independently selected    from halo, (C₁₋₆)alkyl, (C₂₋₆)alkenyl, (C₁₋₆)haloalkyl,    (C₃₋₇)cycloalkyl, —OH, —O(C₁₋₆)alkyl, —SH, —S(C₁₋₆)alkyl, —NH₂,    —NH(C₁₋₆)alkyl and —N((C₁₋₆)alkyl)₂; wherein the (C₁₋₆)alkyl is    optionally substituted with hydroxy, cyano or oxo;-   R⁶ and R⁷ are each independently selected from H, halo, (C₁₋₆)alkyl    and (C₁₋₆)haloalkyl;-   wherein Het is a 4- to 7-membered saturated, unsaturated or aromatic    heterocycle having 1 to 4 heteroatoms each independently selected    from O, N and S, or a 7- to 14-membered saturated, unsaturated or    aromatic heteropolycycle having wherever possible 1 to 5    heteroatoms, each independently selected from O, N and S;-   or a salt or an ester thereof.

Another aspect of the invention provides an isomer, racemate, enantiomeror diasteriomer of compounds of formula (I):

wherein

-   R⁴ is aryl or Het, wherein each of the aryl and Het is optionally    substituted with 1 to 3 substituents each independently selected    from halo, (C₁₋₆)alkyl, (C₂₋₆)alkenyl, (C₁₋₆)haloalkyl,    (C₃₋₇)cycloalkyl, —OH, —O(C₁₋₆)alkyl, —SH, —S(C₁₋₆)alkyl, —NH₂,    —NH(C₁₋₆)alkyl and —N((C₁₋₆)alkyl)₂; wherein the (C₁₋₆)alkyl is    optionally substituted with hydroxy, cyano or oxo; and wherein the    aryl is not monosubstituted at the para position;-   R⁶ and R⁷ are each independently selected from H, halo, (C₁₋₆)alkyl    and (C₁₋₆)haloalkyl;-   wherein Het is a 4- to 7-membered saturated, unsaturated or aromatic    heterocycle having 1 to 4 heteroatoms each independently selected    from O, N and S, or a 7- to 14-membered saturated, unsaturated or    aromatic heteropolycycle having wherever possible 1 to 5    heteroatoms, each independently selected from O, N and S;-   or a salt or an ester thereof.

Another aspect of this invention provides a compound of formula (I), ora pharmaceutically acceptable salt or ester thereof, exhibiting at leastone of the following surprising advantages:

-   -   unexpectedly good activity in a cell-based HIV-1 replication        assay in four of the major integrase variants at residues        124/125 (Thr124/Thr125, Ala124/Thr125, Ala124/Ala125 and        Thr124/Ala125); and/or    -   unexpectedly good activity in a cell-based HIV-1 replication        assay in all six of the major integrase variants at residues        124/125 (Thr124/Thr125, Ala124/Thr125, Ala124/Ala125,        Thr124/Ala125, Asn124/Thr125, and Asn124/Ala125); and/or    -   unexpectedly good pharmacological properties.

Another aspect of this invention provides a compound of formula (I) or apharmaceutically acceptable salt or ester thereof, as a medicament.

Still another aspect of this invention provides a pharmaceuticalcomposition comprising a therapeutically effective amount of a compoundof formula (I) or a pharmaceutically acceptable salt or ester thereof;and one or more pharmaceutically acceptable carriers.

According to an embodiment of this aspect, the pharmaceuticalcomposition according to this invention additionally comprises at leastone other antiviral agent.

The invention also provides the use of a pharmaceutical composition asdescribed hereinabove for the treatment of an HIV infection in a mammalhaving or at risk of having the infection.

A further aspect of the invention involves a method of treating an HIVinfection in a mammal having or at risk of having the infection, themethod comprising administering to the mammal a therapeuticallyeffective amount of a compound of formula (I), a pharmaceuticallyacceptable salt or ester thereof, or a composition thereof as describedhereinabove.

Another aspect of the invention involves a method of treating an HIVinfection in a mammal having or at risk of having the infection, themethod comprising administering to the mammal a therapeuticallyeffective amount of a combination of a compound of formula (I) or apharmaceutically acceptable salt or ester thereof, and at least oneother antiviral agent; or a composition thereof.

Also within the scope of this invention is the use of a compound offormula (I) as described herein, or a pharmaceutically acceptable saltor ester thereof, for the treatment of an HIV infection in a mammalhaving or at risk of having the infection.

Another aspect of this invention provides the use of a compound offormula (I) as described herein, or a pharmaceutically acceptable saltor ester thereof, for the manufacture of a medicament for the treatmentof an HIV infection in a mammal having or at risk of having theinfection.

An additional aspect of this invention refers to an article ofmanufacture comprising a composition effective to treat an HIVinfection; and packaging material comprising a label which indicatesthat the composition can be used to treat infection by HIV; wherein thecomposition comprises a compound of formula (I) according to thisinvention or a pharmaceutically acceptable salt or ester thereof.

Still another aspect of this invention relates to a method of inhibitingthe replication of HIV comprising exposing the virus to an effectiveamount of the compound of formula (I), or a salt or ester thereof, underconditions where replication of HIV is inhibited.

Further included in the scope of the invention is the use of a compoundof formula (I) to inhibit the activity of the HIV integrase enzyme.

Further included in the scope of the invention is the use of a compoundof formula (I), or a salt or ester thereof, to inhibit the replicationof HIV.

DETAILED DESCRIPTION OF THE INVENTION Definitions

As used herein, the following definitions apply unless otherwise noted:

The term “substituent”, as used herein and unless specified otherwise,is intended to mean an atom, radical or group which may be bonded to acarbon atom, a heteroatom or any other atom which may form part of amolecule or fragment thereof, which would otherwise be bonded to atleast one hydrogen atom. Substituents contemplated in the context of aspecific molecule or fragment thereof are those which give rise tochemically stable compounds, such as are recognized by those skilled inthe art.

The term “(C_(1-n))alkyl” as used herein, wherein n is an integer,either alone or in combination with another radical, is intended to meanacyclic, straight or branched chain alkyl radicals containing from 1 ton carbon atoms. “(C₁₋₆)alkyl” includes, but is not limited to, methyl,ethyl, propyl (n-propyl), butyl (n-butyl), 1-methylethyl (iso-propyl),1-methylpropyl (sec-butyl), 2-methylpropyl (iso-butyl),1,1-dimethylethyl (tert-butyl), pentyl and hexyl. The abbreviation Medenotes a methyl group; Et denotes an ethyl group, Pr denotes a propylgroup, iPr denotes a 1-methylethyl group, Bu denotes a butyl group andtBu denotes a 1,1-dimethylethyl group.

The term “(C_(2-n))alkenyl”, as used herein, wherein n is an integer,either alone or in combination with another radical, is intended to meanan unsaturated, acyclic straight or branched chain radical containingtwo to n carbon atoms, at least two of which are bonded to each other bya double bond. Examples of such radicals include, but are not limitedto, ethenyl (vinyl), 1-propenyl, 2-propenyl, and 1-butenyl. Unlessspecified otherwise, the term “(C_(2-n))alkenyl” is understood toencompass individual stereoisomers where possible, including but notlimited to (E) and (Z) isomers, and mixtures thereof. When a(C_(2-n))alkenyl group is substituted, it is understood to besubstituted on any carbon atom thereof which would otherwise bear ahydrogen atom, unless specified otherwise, such that the substitutionwould give rise to a chemically stable compound, such as are recognizedby those skilled in the art.

The term “(C_(3-m))cycloalkyl” as used herein, wherein m is an integer,either alone or in combination with another radical, is intended to meana cycloalkyl substituent containing from 3 to m carbon atoms andincludes, but is not limited to, cyclopropyl, cyclobutyl, cyclopentyl,cyclohexyl and cycloheptyl.

The term “aryl” as used herein, either alone or in combination withanother radical, is intended to mean a carbocyclic aromatic monocyclicgroup containing 6 carbon atoms which may be further fused to a second5- or 6-membered carbocyclic group which may be aromatic, saturated orunsaturated. Aryl includes, but is not limited to, phenyl, indanyl,indenyl, 1-naphthyl, 2-naphthyl, tetrahydronaphthyl and dihydronaphthyl.

The term “carbocycle” as used herein, either alone or in combinationwith another radical, is intended to mean a cyclic compound, eitheraromatic or non-aromatic, saturated or unsaturated, in which all of thering members are carbon atoms. The carbocycle group may be containing 5or 6 carbon atom and may be further fused to a second 5- or 6-memberedcarbocyclic group which may be aromatic, saturated or unsaturated. Thecarbocycle may be substituted. When the carbocycle is substituted, it isunderstood that substituents may be attached to any carbon atom whichwould otherwise bear a hydrogen atom, unless specified otherwise, suchthat the substitution would give rise to a chemically stable compound,such as are recognized by those skilled in the art.

The term “Het” as used herein, either alone or in combination withanother radical, is intended to mean a 4- to 7-membered saturated,unsaturated or aromatic heterocycle having 1 to 4 heteroatoms eachindependently selected from O, N and S, or a 7- to 14-memberedsaturated, unsaturated or aromatic heteropolycycle having whereverpossible 1 to 5 heteroatoms, each independently selected from O, N andS, unless specified otherwise. When a Het group is substituted, it isunderstood that substituents may be attached to any carbon atom orheteroatom thereof which would otherwise bear a hydrogen atom, unlessspecified otherwise, such that the substitution would give rise to achemically stable compound, such as are recognized by those skilled inthe art.

The term “heteroatom” as used herein is intended to mean O, S or N.

The term “heterocycle” as used herein and unless specified otherwise,either alone or in combination with another radical, is intended to meana 3- to 7-membered saturated, unsaturated or aromatic heterocyclecontaining from 1 to 4 heteroatoms each independently selected from O, Nand S; or a monovalent radical derived by removal of a hydrogen atomtherefrom. Examples of such heterocycles include, but are not limitedto, azetidine, pyrrolidine, tetrahydrofuran, tetrahydrothiophene,thiazolidine, oxazolidine, pyrrole, thiophene, furan, pyrazole,imidazole, isoxazole, oxazole, isothiazole, thiazole, triazole,tetrazole, piperidine, piperazine, azepine, diazepine, pyran,1,4-dioxane, 4-morpholine, 4-thiomorpholine, pyridine, pyridine-N-oxide,pyridazine, pyrazine and pyrimidine, and saturated, unsaturated andaromatic derivatives thereof.

The term “heteropolycycle” as used herein and unless specifiedotherwise, either alone or in combination with another radical, isintended to mean a heterocycle as defined above fused to one or moreother cycle, including a carbocycle, a heterocycle or any other cycle;or a monovalent radical derived by removal of a hydrogen atom therefrom.Examples of such heteropolycycles include, but are not limited to,indole, isoindole, benzimidazole, benzothiophene, benzofuran,benzopyran, benzodioxole, benzodioxane, benzothiazole, quinoline,isoquinoline, and naphthyridine, and saturated, unsaturated and aromaticderivatives thereof.

The term “halo” as used herein is intended to mean a halogen substituentselected from fluoro, chloro, bromo or iodo.

The term “(C_(1-n))haloalkyl” as used herein, wherein n is an integer,either alone or in combination with another radical, is intended to meanan alkyl radical having 1 to n carbon atoms as defined above wherein oneor more hydrogen atoms are each replaced by a halo substituent. Examplesof (C_(1-n))haloalkyl include but are not limited to chloromethyl,chloroethyl, dichloroethyl, bromomethyl, bromoethyl, dibromoethyl,fluoromethyl, difluoromethyl, trifluoromethyl, fluoroethyl anddifluoroethyl.

The terms “—O—(C_(1-n))alkyl” or “(C_(1-n))alkoxy” as used hereininterchangeably, wherein n is an integer, either alone or in combinationwith another radical, is intended to mean an oxygen atom further bondedto an alkyl radical having 1 to n carbon atoms as defined above.Examples of —O—(C_(1-n))alkyl include but are not limited to methoxy(CH₃O—), ethoxy (CH₃CH₂O—), propoxy (CH₃CH₂CH₂O—), 1-methylethoxy(iso-propoxy; (CH₃)₂CH—O—) and 1,1-dimethylethoxy (tert-butoxy;(CH₃)₃C—O—). When an —O—(C_(1-n))alkyl radical is substituted, it isunderstood to be substituted on the (C_(1-n))alkyl portion thereof, suchthat the substitution would give rise to a chemically stable compound,such as are recognized by those skilled in the art.

The terms “—S—(C_(1-n))alkyl” or “(C_(1-n))alkylthio” as used hereininterchangeably, wherein n is an integer, either alone or in combinationwith another radical, is intended to mean an sulfur atom further bondedto an alkyl radical having 1 to n carbon atoms as defined above.Examples of —S—(C_(1-n))alkyl include but are not limited to methylthio(CH₃S—), ethylthio (CH₃CH₂S—), propylthio (CH₃CH₂CH₂S—),1-methylethylthio (isopropylthio; (CH₃)₂CH—S—) and 1,1-dimethylethylthio(tert-butylthio; (CH₃)₃C—S—). When —S—(C_(1-n))alkyl radical, or anoxidized derivative thereof, such as an —SO—(C_(1-n))alkyl radical or an—SO₂—(C_(1-n))alkyl radical, is substituted, each is understood to besubstituted on the (C_(1-n))alkyl portion thereof, such that thesubstitution would give rise to a chemically stable compound, such asare recognized by those skilled in the art.

The term “oxo” as used herein is intended to mean an oxygen atomattached to a carbon atom as a substituent by a double bond (═O).

The term “cyano” as used herein is intended to mean an carbon atomattached to a nitrogen atom as a substituent by a triple bond.

The term “functional group equivalent” as used herein is intended tomean an atom or group that may replace another atom or group which hassimilar electronic, hybridization or bonding properties.

The term “protecting group” as used herein is intended to meanprotecting groups that can be used during synthetic transformation,including but not limited to examples which are listed in Greene,“Protective Groups in Organic Chemistry”, John Wiley & Sons, New York(1981), and more recent editions thereof, herein incorporated byreference.

The following designation

is used in sub-formulas to indicate the bond which is connected to therest of the molecule as defined.

The term “salt thereof” as used herein is intended to mean any acidand/or base addition salt of a compound according to the invention,including but not limited to a pharmaceutically acceptable salt thereof.

The term “pharmaceutically acceptable salt” as used herein is intendedto mean a salt of a compound according to the invention which is, withinthe scope of sound medical judgment, suitable for use in contact withthe tissues of humans and lower animals without undue toxicity,irritation, allergic response, and the like, commensurate with areasonable benefit/risk ratio, generally water or oil-soluble ordispersible, and effective for their intended use. The term includespharmaceutically-acceptable acid addition salts andpharmaceutically-acceptable base addition salts. Lists of suitable saltsare found in, for example, S. M. Birge et al., J. Pharm. Sci., 1977, 66,pp. 1-19, herein incorporated by reference.

The term “pharmaceutically-acceptable acid addition salt” as used hereinis intended to mean those salts which retain the biologicaleffectiveness and properties of the free bases and which are notbiologically or otherwise undesirable, formed with inorganic acidsincluding but not limited to hydrochloric acid, hydrobromic acid,sulfuric acid, sulfamic acid, nitric acid, phosphoric acid and the like,and organic acids including but not limited to acetic acid,trifluoroacetic acid, adipic acid, ascorbic acid, aspartic acid,benzenesulfonic acid, benzoic acid, butyric acid, camphoric acid,camphorsulfonic acid, cinnamic acid, citric acid, digluconic acid,ethanesulfonic acid, glutamic acid, glycolic acid, glycerophosphoricacid, hemisulfic acid, hexanoic acid, formic acid, fumaric acid,2-hydroxyethanesulfonic acid (isethionic acid), lactic acid,hydroxymaleic acid, malic acid, malonic acid, mandelic acid,mesitylenesulfonic acid, methanesulfonic acid, naphthalenesulfonic acid,nicotinic acid, 2-naphthalenesulfonic acid, oxalic acid, pamoic acid,pectinic acid, phenylacetic acid, 3-phenylpropionic acid, pivalic acid,propionic acid, pyruvic acid, salicylic acid, stearic acid, succinicacid, sulfanilic acid, tartaric acid, p-toluenesulfonic acid, undecanoicacid and the like.

The term “pharmaceutically-acceptable base addition salt” as used hereinis intended to mean those salts which retain the biologicaleffectiveness and properties of the free acids and which are notbiologically or otherwise undesirable, formed with inorganic basesincluding but not limited to ammonia or the hydroxide, carbonate, orbicarbonate of ammonium or a metal cation such as sodium, potassium,lithium, calcium, magnesium, iron, zinc, copper, manganese, aluminum andthe like. Particularly preferred are the ammonium, potassium, sodium,calcium, and magnesium salts. Salts derived frompharmaceutically-acceptable organic nontoxic bases include but are notlimited to salts of primary, secondary, and tertiary amines, quaternaryamine compounds, substituted amines including naturally occurringsubstituted amines, cyclic amines and basic ion-exchange resins, such asmethylamine, dimethylamine, trimethylamine, ethylamine, diethylamine,triethylamine, isopropylamine, tripropylamine, tributylamine,ethanolamine, diethanolamine, 2-dimethylaminoethanol,2-diethylaminoethanol, dicyclohexylamine, lysine, arginine, histidine,caffeine, hydrabamine, choline, betaine, ethylenediamine, glucosamine,methylglucamine, theobromine, purines, piperazine, piperidine,N-ethylpiperidine, tetramethylammonium compounds, tetraethylammoniumcompounds, pyridine, N,N-dimethylaniline, N-methylpiperidine,N-methylmorpholine, dicyclohexylamine, dibenzylamine,N,N-dibenzylphenethylamine, 1-ephenamine, N,N′-dibenzylethylenediamine,polyamine resins and the like. Particularly preferred organic nontoxicbases are isopropylamine, diethylamine, ethanolamine, trimethylamine,dicyclohexylamine, choline, and caffeine.

The term “ester thereof” as used herein is intended to mean any ester ofa compound according to the invention in which any of the —COOHsubstituents of the molecule is replaced by a —COOR substituent, inwhich the R moiety of the ester is any carbon-containing group whichforms a stable ester moiety, including but not limited to alkyl,alkenyl, alkynyl, cycloalkyl, cycloalkylalkyl, aryl, arylalkyl,heterocyclyl, heterocyclylalkyl, each of which being optionally furthersubstituted. The term “ester thereof” includes but is not limited topharmaceutically acceptable esters thereof.

The term “pharmaceutically acceptable ester” as used herein is intendedto mean esters of the compound according to the invention in which anyof the COOH substituents of the molecule are replaced by a —COORsubstituent, in which the R moiety of the ester is selected from alkyl(including, but not limited to, methyl, ethyl, propyl, 1-methylethyl,1,1-dimethylethyl, butyl); alkoxyalkyl (including, but not limited tomethoxymethyl); acyloxyalkyl (including, but not limited toacetoxymethyl); arylalkyl (including, but not limited to, benzyl);aryloxyalkyl (including, but not limited to, phenoxymethyl); and aryl(including, but not limited to phenyl) optionally substituted withhalogen, (C₁₋₄)alkyl or (C₁₋₄)alkoxy. Other suitable esters can be foundin Design of Prodrugs, Bundgaard, H. Ed. Elsevier (1985), hereinincorporated by reference. Such pharmaceutically acceptable esters areusually hydrolyzed in vivo when injected into a mammal and transformedinto the acid form of the compound according to the invention. Withregard to the esters described above, unless otherwise specified, anyalkyl moiety present preferably contains 1 to 16 carbon atoms, morepreferably 1 to 6 carbon atoms. Any aryl moiety present in such esterspreferably comprises a phenyl group. In particular the esters may be a(C₁₋₁₆)alkyl ester, an unsubstituted benzyl ester or a benzyl estersubstituted with at least one halogen, (C₁₋₆)alkyl, (C₁₋₆)alkoxy, nitroor trifluoromethyl.

The term “mammal” as used herein is intended to encompass humans, aswell as non-human mammals which are susceptible to infection by HIV.Non-human mammals include but are not limited to domestic animals, suchas cows, pigs, horses, dogs, cats, rabbits, rats and mice, andnon-domestic animals.

The term “treatment” as used herein is intended to mean theadministration of a compound or composition according to the presentinvention to alleviate or eliminate symptoms of HIV infection and/or toreduce viral load in a patient. The term “treatment” also encompassesthe administration of a compound or composition according to the presentinvention post-exposure of the individual to the virus but before theappearance of symptoms of the disease, and/or prior to the detection ofthe virus in the blood, to prevent the appearance of symptoms of thedisease and/or to prevent the virus from reaching detectible levels inthe blood, and the administration of a compound or composition accordingto the present invention to prevent perinatal transmission of HIV frommother to baby, by administration to the mother before giving birth andto the child within the first days of life.

The term “antiviral agent” as used herein is intended to mean an agentthat is effective to inhibit the formation and/or replication of a virusin a mammal, including but not limited to agents that interfere witheither host or viral mechanisms necessary for the formation and/orreplication of a virus in a mammal.

The term “inhibitor of HIV replication” as used herein is intended tomean an agent capable of reducing or eliminating the ability of HIV toreplicate in a host cell, whether in vitro, ex vivo or in vivo.

The term “HIV integrase” or “integrase”, used herein interchangeably,means the integrase enzyme encoded by the human immunodeficiency virustype 1.

The term “therapeutically effective amount” means an amount of acompound according to the invention, which when administered to apatient in need thereof, is sufficient to effect treatment fordisease-states, conditions, or disorders for which the compounds haveutility. Such an amount would be sufficient to elicit the biological ormedical response of a tissue system, or patient that is sought by aresearcher or clinician. The amount of a compound according to theinvention which constitutes a therapeutically effective amount will varydepending on such factors as the compound and its biological activity,the composition used for administration, the time of administration, theroute of administration, the rate of excretion of the compound, theduration of the treatment, the type of disease-state or disorder beingtreated and its severity, drugs used in combination with orcoincidentally with the compounds of the invention, and the age, bodyweight, general health, sex and diet of the patient. Such atherapeutically effective amount can be determined routinely by one ofordinary skill in the art having regard to their own knowledge, thestate of the art, and this disclosure.

PREFERRED EMBODIMENTS

In the following preferred embodiments, groups and substituents of thecompounds of formula (I):

according to this invention are described in detail.

-   R⁴:-   R⁴-A: In one embodiment, R⁴ is aryl or Het optionally substituted    with 1 to 3 substituents each independently selected from halo,    (C₁₋₆)alkyl, (C₂₋₆)alkenyl, (C₁₋₆)haloalkyl, (C₃₋₇)cycloalkyl, —OH,    —O(C₁₋₆)alkyl, —SH, —S(C₁₋₆)alkyl, —NH₂, —NH(C₁₋₆)alkyl and    —N((C₁₋₆)alkyl)₂; wherein the (C₁₋₆)alkyl is optionally substituted    with hydroxy, cyano or oxo.-   R⁴-B: In another embodiment, R⁴ is Het optionally substituted with 1    to 3 substituents each independently selected from halo,    (C₁₋₆)alkyl, (C₂₋₆)alkenyl, (C₁₋₆)haloalkyl, (C₃₋₇)cycloalkyl, —OH,    —O(C₁₋₆)alkyl, —SH, —S(C₁₋₆)alkyl, —NH₂, —NH(C₁₋₆)alkyl and    —N((C₁₋₆)alkyl)₂.-   R⁴-C: In another embodiment, R⁴ is naphthyl or phenyl, optionally    substituted with 1 to 3 substituents each independently selected    from halo, (C₁₋₄)alkyl, (C₁₋₄)haloalkyl, —O(C₁₋₄)alkyl, —NH₂,    —NH(C₁₋₆)alkyl and —N((C₁₋₆)alkyl)₂.-   R⁴-D: In another embodiment, R⁴ is phenyl optionally substituted    with 1 to 3 substituents each independently selected from halo,    (C₁₋₄)alkyl, (C₁₋₄)haloalkyl, —O(C₁₋₄)alkyl, —NH₂, —NH(C₁₋₆)alkyl    and —N((C₁₋₆)alkyl)₂.-   R⁴-E: In one embodiment, R⁴ is Het optionally substituted with 1 to    2 substituents each independently selected from halo, (C₁₋₃)alkyl    and O—(C₁₋₃)alkyl.-   R⁴-F: In one embodiment, R⁴ is Het optionally substituted with 1 to    2 substituents each independently selected from Cl, F, CH₃ and    CH₂CH₃ wherein said Het is defined as a 7- to 14-membered saturated,    unsaturated or aromatic heteropolycycle having wherever possible 1    to 2 heteroatoms, each independently selected from O, N and S.-   R⁴-G: In one embodiment, R⁴ is Het optionally substituted with 1 to    2 substituents each independently selected from halo, (C₁₋₃)alkyl    and O—(C₁₋₃)alkyl, wherein said Het is defined as a 9- or    10-membered saturated, unsaturated or aromatic heteropolycycle    having wherever possible 1 to 2 heteroatoms, each independently    selected from O, N and S.-   R⁴-H: In another embodiment, R⁴ is phenyl optionally substituted    with 1 to 3 substituents each independently selected from halo,    (C₁₋₄)alkyl, (C₁₋₄)haloalkyl, —O(C₁₋₄)alkyl, —NH₂, —NH(C₁₋₆)alkyl    and —N((C₁₋₆)alkyl)₂ or R⁴ is Het optionally substituted with 1 to 3    substituents each independently selected from halo, (C₁₋₄)alkyl and    O—(C₁₋₄)alkyl, wherein said Het is defined as a 7- to 14-membered    saturated, unsaturated or aromatic heteropolycycle having wherever    possible 1 to 2 heteroatoms, each independently selected from O, N    and S.-   R⁴-I: In another embodiment, R⁴ is phenyl optionally substituted    with 1 to 3 substituents each independently selected from F, Cl, Br,    —CH₃, —CH(CH₃)₂, CH₂F, —CH₂CH₂F, —OCH₃ and —NH₂ or R⁴ is Het    optionally substituted with 1 to 3 substituents each independently    selected from Cl, F, CH₃, CH₂CH₃ and OCH₃, wherein said Het is    defined as a 7- or 14-membered saturated, unsaturated or aromatic    heteropolycycle having wherever possible 1 to 3 heteroatoms, each    independently selected from O, N and S.-   R⁴-J: In another embodiment, R⁴ is selected from:

-   -   being optionally substituted 1 to 3 times with halo, (C₁₋₃)alkyl        and O—(C₁₋₃)alkyl.

-   R⁴-K: In another embodiment, R⁴ is selected from:

-   -   being optionally substituted 1 to 2 times with halo, (C₁₋₃)alkyl        and O—(C₁₋₃)alkyl.

-   R⁴-L: In another embodiment, R⁴ is selected from:

-   -   being optionally substituted 1 to 2 times with halo, (C₁₋₃)alkyl        and O—(C₁₋₃)alkyl.

-   R⁴-M: In another embodiment, R⁴ is selected from:

-   -   being optionally substituted 1 to 2 times with halo, (C₁₋₃)alkyl        and O—(C₁₋₃)alkyl.

-   R⁴-N: In another embodiment, R⁴ is selected from:

-   R⁴-O: In another embodiment, R⁴ is selected from:

One skilled in the art will recognize that when the R⁴ substituent isnot symmetrically substituted about the axis of rotation of the bondattaching R⁴ to Core, rotational isomers or atropisomers are possible.Compounds of the invention in which the R⁴ substituent is notsymmetrically substituted about the axis of rotation of the bondattaching R⁴ to Core and in which the carbon atom bonded to the —COOHand R³ substituents is chiral, as described above, will have two chiralcenters, a chiral carbon atom and a rotational axis of asymmetry, andthus the atropisomers will exist as diastereomers. However, individualdiastereomeric atropisomers may or may not be detectable and/orseparable, depending upon the relative amounts of each atropisomerformed during synthesis, present at equilibrium, and the degree ofsteric hindrance to rotation about the C-4 chiral axis, and therefore,the rate at which interconversion between these atropoisomers occurs.Once separated, individual atropoisomers may be very stable orinterconvert, rapidly or slowly, with each other to form an equilibriummixture of atropoisomers.

-   R⁴-P: In another embodiment, R⁴ is selected from:

-   R⁴-Q: In another embodiment, R⁴ is selected from:

-   R⁴-R: In another embodiment, R⁴ is aryl or Het, wherein each of the    aryl and Het is optionally substituted with 1 to 3 substituents each    independently selected from halo, (C₁₋₆)alkyl, (C₂₋₆)alkenyl,    (C₁₋₆)haloalkyl, (C₃₋₇)cycloalkyl, —OH, —O(C₁₋₆)alkyl, —SH,    —S(C₁₋₆)alkyl, —NH₂, —NH(C₁₋₆)alkyl and —N((C₁₋₆)alkyl)₂; wherein    the (C₁₋₆)alkyl is optionally substituted with hydroxy, cyano or    oxo; and wherein the aryl is not monosubstituted at the para    position;

Any and each individual definition of R⁴ as set out herein may becombined with any and each individual definition of R⁶ and R⁷ as set outherein.

-   R⁶:-   R⁶-A: In one embodiment, R⁶ is H, halo, (C₁₋₆)alkyl or    (C₁₋₆)haloalkyl.-   R⁶-B: In another embodiment, R⁶ is H, halo or (C₁₋₃)alkyl.-   R⁶-C: In another embodiment, R⁶ is H, F, Cl or (C₁₋₂)alkyl.-   R⁶-D: In another embodiment, R⁶ is H, F, C₁ or CH₃.-   R⁶-E: In another embodiment, R⁶ is H, CH₃ or CH₂CH₃.-   R⁶-F: In another embodiment, R⁶ is H or CH₃.-   R⁶-G: In another embodiment, R⁶ is H.

Any and each individual definition of R⁶ as set out herein may becombined with any and each individual definition of R⁴ and R⁷ as set outherein.

-   R⁷:-   R⁷-A: In one embodiment, R⁷ is H, halo, (C₁₋₆)alkyl or    (C₁₋₆)haloalkyl.-   R⁷-B: In another embodiment, R⁷ is H, halo or (C₁₋₃)alkyl.-   R⁷-C: In another embodiment, R⁷ is H, F, Cl or (C₁₋₂)alkyl.-   R⁷-D: In another embodiment, R⁷ is H, F, C₁ or CH₃.-   R⁷-E: In one embodiment, R⁷ is H, F or CH₃.-   R⁷-F: In one embodiment, R⁷ is H or CH₃.-   R⁷-G: In another embodiment, R⁷ is H.

Any and each individual definition of R⁷ as set out herein may becombined with any and each individual definition of R⁴ and R⁶ as set outherein.

Examples of preferred subgeneric embodiments of the present inventionare set forth in the following table, wherein each substituent group ofeach embodiment is defined according to the definitions set forth above:

Embodiment R⁴ R⁶ R⁷ E-1 R⁴-A R⁶-B R⁷-C E-2 R⁴-A R⁶-D R⁷-C E-3 R⁴-A R⁶-ER⁷-F E-4 R⁴-A R⁶-F R⁷-A E-5 R⁴-A R⁶-E R⁷-E E-6 R⁴-B R⁶-E R⁷-E E-7 R⁴-BR⁶-B R⁷-F E-8 R⁴-B R⁶-F R⁷-A E-9 R⁴-C R⁶-A R⁷-A E-10 R⁴-C R⁶-D R⁷-A E-11R⁴-C R⁶-F R⁷-D E-12 R⁴-D R⁶-E R⁷-F E-13 R⁴-D R⁶-D R⁷-F E-14 R⁴-D R⁶-GR⁷-F E-15 R⁴-D R⁶-E R⁷-G E-16 R⁴-D R⁶-D R⁷-C E-17 R⁴-D R⁶-E R⁷-C E-18R⁴-E R⁶-C R⁷-A E-19 R⁴-E R⁶-B R⁷-C E-20 R⁴-E R⁶-B R⁷-B E-21 R⁴-E R⁶-BR⁷-E E-22 R⁴-E R⁶-E R⁷-C E-23 R⁴-F R⁶-D R⁷-A E-24 R⁴-F R⁶-E R⁷-C E-25R⁴-F R⁶-C R⁷-B E-26 R⁴-F R⁶-B R⁷-E E-27 R⁴-G R⁶-A R⁷-D E-28 R⁴-G R⁶-CR⁷-B E-29 R⁴-G R⁶-B R⁷-A E-30 R⁴-G R⁶-B R⁷-G E-31 R⁴-G R⁶-A R⁷-F E-32R⁴-G R⁶-A R⁷-E E-33 R⁴-G R⁶-C R⁷-D E-34 R⁴-H R⁶-C R⁷-E E-35 R⁴-H R⁶-CR⁷-F E-36 R⁴-H R⁶-D R⁷-E E-37 R⁴-H R⁶-D R⁷-F E-38 R⁴-H R⁶-E R⁷-E E-39R⁴-H R⁶-E R⁷-F E-40 R⁴-H R⁶-G R⁷-B E-41 R⁴-H R⁶-C R⁷-F E-42 R⁴-H R⁶-FR⁷-C E-43 R⁴-H R⁶-G R⁷-E E-44 R⁴-H R⁶-D R⁷-E E-45 R⁴-H R⁶-D R⁷-A E-46R⁴-H R⁶-A R⁷-B E-47 R⁴-I R⁶-C R⁷-E E-48 R⁴-I R⁶-C R⁷-F E-49 R⁴-I R⁶-DR⁷-E E-50 R⁴-I R⁶-D R⁷-F E-51 R⁴-I R⁶-E R⁷-E E-52 R⁴-I R⁶-E R⁷-F E-53R⁴-I R⁶-B R⁷-C E-54 R⁴-I R⁶-A R⁷-G E-55 R⁴-I R⁶-B R⁷-C E-56 R⁴-J R⁶-CR⁷-E E-57 R⁴-J R⁶-C R⁷-F E-58 R⁴-J R⁶-D R⁷-E E-59 R⁴-J R⁶-D R⁷-F E-60R⁴-J R⁶-E R⁷-E E-61 R⁴-J R⁶-E R⁷-F E-62 R⁴-J R⁶-F R⁷-D E-63 R⁴-J R⁶-AR⁷-A E-64 R⁴-J R⁶-F R⁷-G E-65 R⁴-J R⁶-G R⁷-F E-66 R⁴-J R⁶-C R⁷-F E-67R⁴-J R⁶-D R⁷-G E-68 R⁴-J R⁶-G R⁷-E E-69 R⁴-K R⁶-C R⁷-E E-70 R⁴-K R⁶-CR⁷-F E-71 R⁴-K R⁶-D R⁷-E E-72 R⁴-K R⁶-D R⁷-F E-73 R⁴-K R⁶-E R⁷-E E-74R⁴-K R⁶-E R⁷-F E-75 R⁴-K R⁶-G R⁷-A E-76 R⁴-K R⁶-B R⁷-C E-77 R⁴-K R⁶-GR⁷-E E-78 R⁴-L R⁶-C R⁷-F E-79 R⁴-L R⁶-D R⁷-E E-80 R⁴-L R⁶-D R⁷-F E-81R⁴-L R⁶-E R⁷-E E-82 R⁴-L R⁶-E R⁷-F E-83 R⁴-L R⁶-G R⁷-A E-84 R⁴-L R⁶-BR⁷-C E-85 R⁴-L R⁶-G R⁷-E E-86 R⁴-L R⁶-C R⁷-F E-87 R⁴-L R⁶-F R⁷-F E-88R⁴-L R⁶-F R⁷-G E-89 R⁴-L R⁶-A R⁷-C E-90 R⁴-L R⁶-D R⁷-A E-91 R⁴-M R⁶-CR⁷-F E-92 R⁴-M R⁶-D R⁷-E E-93 R⁴-M R⁶-D R⁷-F E-94 R⁴-M R⁶-E R⁷-E E-95R⁴-M R⁶-E R⁷-F E-96 R⁴-M R⁶-G R⁷-A E-97 R⁴-M R⁶-B R⁷-C E-98 R⁴-M R⁶-GR⁷-E E-99 R⁴-M R⁶-C R⁷-F E-100 R⁴-M R⁶-F R⁷-F E-101 R⁴-M R⁶-F R⁷-G E-102R⁴-M R⁶-A R⁷-D E-103 R⁴-M R⁶-C R⁷-B E-104 R⁴-N R⁶-C R⁷-E E-105 R⁴-N R⁶-CR⁷-F E-106 R⁴-N R⁶-D R⁷-E E-107 R⁴-N R⁶-D R⁷-F E-108 R⁴-N R⁶-E R⁷-EE-109 R⁴-N R⁶-E R⁷-F E-110 R⁴-N R⁶-F R⁷-G E-111 R⁴-N R⁶-B R⁷-B E-112R⁴-N R⁶-G R⁷-A E-113 R⁴-N R⁶-A R⁷-G E-114 R⁴-O R⁶-C R⁷-E E-115 R⁴-O R⁶-CR⁷-F E-116 R⁴-O R⁶-D R⁷-E E-117 R⁴-O R⁶-D R⁷-F E-118 R⁴-O R⁶-E R⁷-EE-119 R⁴-O R⁶-E R⁷-F E-120 R⁴-O R⁶-B R⁷-F E-121 R⁴-O R⁶-G R⁷-B E-122R⁴-O R⁶-F R⁷-A E-123 R⁴-P R⁶-B R⁷-A E-124 R⁴-P R⁶-C R⁷-B E-125 R⁴-P R⁶-DR⁷-C E-126 R⁴-P R⁶-E R⁷-D E-127 R⁴-P R⁶-F R⁷-E E-128 R⁴-P R⁶-G R⁷-FE-129 R⁴-P R⁶-A R⁷-G E-130 R⁴-P R⁶-A R⁷-A E-131 R⁴-P R⁶-B R⁷-E E-132R⁴-P R⁶-C R⁷-F E-133 R⁴-P R⁶-D R⁷-G E-134 R⁴-P R⁶-E R⁷-E E-135 R⁴-P R⁶-FR⁷-F E-136 R⁴-P R⁶-G R⁷-G E-137 R⁴-Q R⁶-B R⁷-A E-138 R⁴-Q R⁶-C R⁷-BE-139 R⁴-Q R⁶-D R⁷-C E-140 R⁴-Q R⁶-E R⁷-D E-141 R⁴-Q R⁶-F R⁷-E E-142R⁴-Q R⁶-G R⁷-F E-143 R⁴-Q R⁶-A R⁷-G E-144 R⁴-Q R⁶-A R⁷-A E-145 R⁴-Q R⁶-BR⁷-E E-146 R⁴-Q R⁶-C R⁷-F E-147 R⁴-Q R⁶-D R⁷-G E-148 R⁴-Q R⁶-E R⁷-EE-149 R⁴-Q R⁶-F R⁷-F E-150 R⁴-Q R⁶-G R⁷-G

Examples of most preferred compounds according to this invention areeach single compound listed in the following Table 1.

In general, all tautomeric and isomeric forms and mixtures thereof, forexample, individual tautomers, geometric isomers, stereoisomers,atropisomers, enantiomers, diastereomers, racemates, racemic ornon-racemic mixtures of stereoisomers, mixtures of diastereomers, ormixtures of any of the foregoing forms of a chemical structure orcompound is intended, unless the specific stereochemistry or isomericform is specifically indicated in the compound name or structure.

It is well-known in the art that the biological and pharmacologicalactivity of a compound is sensitive to the stereochemistry of thecompound. Thus, for example, enantiomers often exhibit strikinglydifferent biological activity including differences in pharmacokineticproperties, including metabolism, protein binding, and the like, andpharmacological properties, including the type of activity displayed,the degree of activity, toxicity, and the like. Thus, one skilled in theart will appreciate that one enantiomer may be more active or mayexhibit beneficial effects when enriched relative to the otherenantiomer or when separated from the other enantiomer. Additionally,one skilled in the art would know how to separate, enrich, orselectively prepare the enantiomers of the compounds of the presentinvention from this disclosure and the knowledge in the art.

Preparation of pure stereoisomers, e.g. enantiomers and diastereomers,or mixtures of desired enantiomeric excess (ee) or enantiomeric purity,are accomplished by one or more of the many methods of (a) separation orresolution of enantiomers, or (b) enantioselective synthesis known tothose of skill in the art, or a combination thereof. These resolutionmethods generally rely on chiral recognition and include, for example,chromatography using chiral stationary phases, enantioselectivehost-guest complexation, resolution or synthesis using chiralauxiliaries, enantioselective synthesis, enzymatic and nonenzymatickinetic resolution, or spontaneous enantioselective crystallization.Such methods are disclosed generally in Chiral Separation Techniques: APractical Approach (2nd Ed.), G. Subramanian (ed.), Wiley-VCH, 2000; T.E. Beesley and R. P. W. Scott, Chiral Chromatography, John Wiley & Sons,1999; and Satinder Ahuja, Chiral Separations by Chromatography, Am.Chem. Soc., 2000, herein incorporated by reference. Furthermore, thereare equally well-known methods for the quantitation of enantiomericexcess or purity, for example, GC, HPLC, CE, or NMR, and assignment ofabsolute configuration and conformation, for example, CD, ORD, X-raycrystallography, or NMR.

Pharmaceutical Composition

Compounds of the present invention may be administered to a mammal inneed of treatment for HIV infection as a pharmaceutical compositioncomprising a therapeutically effective amount of a compound according tothe invention or a pharmaceutically acceptable salt or ester thereof;and one or more conventional non-toxic pharmaceutically-acceptablecarriers, adjuvants or vehicles. The specific formulation of thecomposition is determined by the solubility and chemical nature of thecompound, the chosen route of administration and standard pharmaceuticalpractice. The pharmaceutical composition according to the presentinvention may be administered orally or systemically.

When one enantiomer of a chiral active ingredient has a differentbiological activity than the other, it is contemplated that thepharmaceutical composition according to the invention may comprise aracemic mixture of the active ingredient, a mixture enriched in oneenantiomer of the active ingredient or a pure enantiomer of the activeingredient. The mixture enriched in one enantiomer of the activeingredient is contemplated to contain from more than 50% to about 100%of one enantiomer of the active ingredient and from about 0% to lessthan 50% of the other enantiomer of the active ingredient. Preferably,when the composition comprises a mixture enriched in one enantiomer ofthe active ingredient or a pure enantiomer of the active ingredient, thecomposition comprises from more than 50% to about 100% of, or only, themore physiologically active enantiomer and/or the less toxic enantiomer.It is well known that one enantiomer of an active ingredient may be themore physiologically active for one therapeutic indication while theother enantiomer of the active ingredient may be the morephysiologically active for a different therapeutic indication; thereforethe preferred enantiomeric makeup of the pharmaceutical composition maydiffer for use of the composition in treating different therapeuticindications.

For oral administration, the compound, or a pharmaceutically acceptablesalt or ester thereof, can be formulated in any orally acceptable dosageform including but not limited to aqueous suspensions and solutions,capsules, powders, syrups, elixirs or tablets. For systemicadministration, including but not limited to administration bysubcutaneous, intracutaneous, intravenous, intramuscular,intra-articular, intrasynovial, intrasternal, intrathecal, andintralesional injection or infusion techniques, it is preferred to use asolution of the compound, or a pharmaceutically acceptable salt or esterthereof, in a pharmaceutically acceptable sterile aqueous vehicle.

Pharmaceutically acceptable carriers, adjuvants, vehicles, diluents,excipients and additives as well as methods of formulatingpharmaceutical compositions for various modes of administration arewell-known to those of skill in the art and are described inpharmaceutical texts such as Remington: The Science and Practice ofPharmacy, 21st Edition, Lippincott Williams & Wilkins, 2005; and L. V.Allen, N. G. Popovish and H. C. Ansel, Pharmaceutical Dosage Forms andDrug Delivery Systems, 8th ed., Lippincott Williams & Wilkins, 2004,herein incorporated by reference.

The dosage administered will vary depending upon known factors,including but not limited to the activity and pharmacodynamiccharacteristics of the specific compound employed and its mode, time androute of administration; the age, diet, gender, body weight and generalhealth status of the recipient; the nature and extent of the symptoms;the severity and course of the infection; the kind of concurrenttreatment; the frequency of treatment; the effect desired; and thejudgment of the treating physician. In general, the compound is mostdesirably administered at a dosage level that will generally affordantivirally effective results without causing any harmful or deleteriousside effects.

A daily dosage of active ingredient can be expected to be about 0.001 toabout 100 milligrams per kilogram of body weight, with the preferreddose being about 0.01 to about 50 mg/kg. Typically, the pharmaceuticalcomposition of this invention will be administered from about 1 to about5 times per day or alternatively, as a continuous infusion. Suchadministration can be used as a chronic or acute therapy. The amount ofactive ingredient that may be combined with the carrier materials toproduce a single dosage form will vary depending upon the host treatedand the particular mode of administration. A typical preparation willcontain from about 5% to about 95% active compound (w/w). Preferably,such preparations contain from about 20% to about 80% active compound.

Therefore, according to one embodiment, the pharmaceutical compositionaccording to the invention comprises a racemic mixture of the compoundof formula (I), or a pharmaceutically acceptable salt or ester thereof.

An alternative embodiment provides a pharmaceutical compositioncomprising a mixture enriched in one enantiomer of the compound offormula (I), or a pharmaceutically acceptable salt or ester thereof.

A further embodiment provides a pharmaceutical composition comprising apure enantiomer of the compound of formula (I), or a pharmaceuticallyacceptable salt or ester thereof.

Combination Therapy

Combination therapy is contemplated wherein a compound according to theinvention, or a pharmaceutically acceptable salt or ester thereof, isco-administered with at least one additional antiviral agent. Theadditional agents may be combined with compounds of this invention tocreate a single dosage form. Alternatively these additional agents maybe separately administered, concurrently or sequentially, as part of amultiple dosage form.

When the pharmaceutical composition of this invention comprises acombination of a compound according to the invention, or apharmaceutically acceptable salt or ester thereof, and one or moreadditional antiviral agent, both the compound and the additional agentshould be present at dosage levels of between about 10 to 100%, and morepreferably between about 10 and 80% of the dosage normally administeredin a monotherapy regimen. In the case of a synergistic interactionbetween the compound of the invention and the additional antiviral agentor agents, the dosage of any or all of the active agents in thecombination may be reduced compared to the dosage normally administeredin a monotherapy regimen.

Antiviral agents contemplated for use in such combination therapyinclude agents (compounds or biologicals) that are effective to inhibitthe formation and/or replication of a virus in a mammal, including butnot limited to agents that interfere with either host or viralmechanisms necessary for the formation and/or replication of a virus ina mammal. Such agents can be selected from:

-   -   NRTIs (nucleoside or nucleotide reverse transcriptase        inhibitors) including but not limited to zidovudine (AZT),        didanosine (ddI), zalcitabine (ddC), stavudine (d4T), lamivudine        (3TC), emtricitabine, abacavir succinate, elvucitabine, adefovir        dipivoxil, lobucavir (BMS-180194) lodenosine (FddA) and        tenofovir including tenofovir disoproxil and tenofovir        disoproxil fumarate salt, COMBIVIR™ (contains 3TC and AZT),        TRIZIVIR™ (contains abacavir, 3TC and AZT), TRUVADA™ (contains        tenofovir and emtricitabine), EPZICOM™ (contains abacavir and        3TC);    -   NNRTIs (non-nucleoside reverse transcriptase inhibitors)        including but not limited to nevirapine, delaviradine,        efavirenz, etravirine and rilpivirine;    -   protease inhibitors including but not limited to ritonavir,        tipranavir, saquinavir, nelfinavir, indinavir, amprenavir,        fosamprenavir, atazanavir, lopinavir, darunavir (TMC-114),        lasinavir and brecanavir (VX-385);    -   entry inhibitors including but not limited to        -   CCR5 antagonists (including but not limited to maraviroc,            vicriviroc, INCB9471 and TAK-652),        -   CXCR4 antagonists (including but not limited to AMD-11070),        -   fusion inhibitors (including but not limited to enfuvirtide            (T-20), TR1-1144 and TR1-999) and        -   others (including but not limited to BMS-488043);    -   integrase inhibitors (including but not limited to raltegravir        (MK-0518), BMS-707035 and elvitegravir (GS 9137));    -   TAT inhibitors;    -   maturation inhibitors (including but not limited to berivimat        (PA-457));    -   immunomodulating agents (including but not limited to        levamisole); and    -   other antiviral agents including hydroxyurea, ribavirin, IL-2,        IL-12 and pensafuside.

Furthermore, a compound according to the invention can be used with atleast one other compound according to the invention or with one or moreantifungal or antibacterial agents (including but not limited tofluconazole).

Therefore, according to one embodiment, the pharmaceutical compositionof this invention additionally comprises one or more antiviral agents.

A further embodiment provides the pharmaceutical composition of thisinvention wherein the one or more antiviral agent comprises at least oneNNRTI.

According to another embodiment of the pharmaceutical composition ofthis invention, the one or more antiviral agent comprises at least oneNRTI.

According to yet another embodiment of the pharmaceutical composition ofthis invention, the one or more antiviral agent comprises at least oneprotease inhibitor.

According to still another embodiment of the pharmaceutical compositionof this invention, the one or more antiviral agent comprises at leastone entry inhibitor.

According to a further embodiment of the pharmaceutical composition ofthis invention, the one or more antiviral agent comprises at least oneintegrase inhibitor.

A compound according to the present invention may also be used as alaboratory reagent or a research reagent. For example, a compound of thepresent invention may be used as positive control to validate assays,including but not limited to surrogate cell-based assays and in vitro orin vivo viral replication assays.

Furthermore, a compound according to the present invention may be usedto treat or prevent viral contamination of materials and thereforereduce the risk of viral infection of laboratory or medical personnel orpatients who come in contact with such materials (e.g. blood, tissue,surgical instruments and garments, laboratory instruments and garments,and blood collection apparatuses and materials).

Derivatives Comprising a Detectable Label

Another aspect of the invention provides a derivative of a compound offormula (I), the derivative comprising a detectable label. Such a labelallows recognition either directly or indirectly of the derivative suchthat it can be detected, measured or quantified. The detectable labelmay itself be detectable, measurable or quantifiable, or it may interactwith one or more other moities which themselves comprise one or moredetectable labels, so that the interaction therebetween allows thederivative to be detected, measured or quantified.

Such derivatives may be used as probes to study HIV replication,including but not limited to study of the mechanism of action of viraland host proteins involved in HIV replication, study of conformationalchanges undergone by such viral and host proteins under variousconditions and study of interactions with entities which bind to orotherwise interact with these viral and host proteins. Derivativesaccording to this aspect of the invention may be used in assays toidentify compounds which interact with viral and host proteins, theassays including but not limited to displacement assays which measurethe extent to which the derivative is displaced from interacting withthe viral and host proteins. A preferred used of derivatives accordingto this aspect of the invention is in displacement assays to identifyHIV integrase inhibitors. Such derivatives may also be used to formcovalent or non-covalent interactions with the viral and host proteinsor to identify residues of the viral and host proteins which interactwith the compounds of the invention.

Detectable labels contemplated for use with derivatives of the compoundsof the invention include, but are not limited to, fluorescent labels,chemiluminescent labels, chromophores, antibodies, enzymatic markers,radioactive isotopes, affinity tags and photoreactive groups.

A fluorescent label is a label which fluoresces, emitting light of onewavelength upon absorption of light of a different wavelength.Fluorescent labels include but are not limited to fluorescein; TexasRed; aminomethylcoumarin; rhodamine dyes, including but not limited totetramethylrhodamine (TAMRA); Alexa dyes including but not limited toAlexa Fluor® 555; cyanine dyes including but not limited to Cy3;europium or lanthanide series based fluorescent molecules; and the like.

A chemiluminescent label is a label which can undergo a chemicalreaction which produces light. Chemiluminescent labels include but arenot limited to luminol, luciferin, lucigenin, and the like.

A chromophore is a label which selectively absorbs certain wavelengthsof visible light while transmitting or reflecting others, therebycausing the compounds which contain the chromophore to appear colored.Chromophores include but are not limited to natural and synthetic dyes.

An antibody is a protein produced by a mammalian immune system inresponse to a specific antigen, which binds specifically to thatantigen. Antibodies contemplated for use as detectable labels accordingto the invention include but are not limited to antibodies against thefollowing: polyhistidine tags, glutathione-S-transferase (GST),hemagglutinin (HA), FLAG® epitope tags, Myc tag, maltose binding protein(MBP), green fluorescent protein (GFP) and the like.

An enzymatic marker is an enzyme whose presence may be detected by meansof an assay specific to the catalytic activity of the enzyme. Enzymaticmarkers contemplated for use as detectable labels according to theinvention include but are not limited to luciferase, horseradishperoxidase (HRP), β-galactosidase and the like.

A radioactive isotope is an isotope of an atom which produces radiationupon radioactive decay. Radioactive isotopes include but are not limitedto ¹⁴C, ³H, ³¹P, ¹²¹I, ¹²⁵I and the like.

An affinity tag is a label which has a strong affinity for anothermoiety, designated herein as a binding partner. Such an affinity tag canbe used to form a complex with the binding partner so that the complexmay be selectively detected or separated from a mixture. Affinity tagsinclude but are not limited to biotin or a derivative thereof, ahistidine polypeptide, a polyarginine, an amylose sugar moiety or adefined epitope recognizable by a specific antibody; suitable epitopesinclude but are not limited to glutathione-S-transferase (GST),hemagglutinin (HA), FLAG® epitope tags, Myc tag, maltose binding protein(MBP), green fluorescent protein (GFP) and the like.

Furthermore, compounds of the invention used as probes may be labelledwith a photoreactive group which is transformed, upon activation bylight, from an inert group to a reactive species, such as a freeradical. Such a group may be used to activate the derivative so that itcan form a covalent bond with one or more residues of a viral or hostprotein. Photoreactive groups include but are not limited tophotoaffinity labels such as benzophenone and azide groups.

Methodology and Synthesis

The synthesis of compounds of formula (I) according to this invention isconveniently accomplished following the general procedure outlined inthe schemes below wherein R⁴, R⁶ and R⁷ are as defined herein. Furtherinstruction is provided to one skilled in the art by the specificexamples set out herein below.

wherein R⁴², R⁴³, R⁴⁴, R⁴⁵ and R⁴⁶ may either be substituents on thephenyl moiety or (R⁴² and R⁴³), (R⁴³ and R⁴⁴), (R⁴⁴ and R⁴⁵) or (R⁴⁵ andR⁴⁶) may be linked so to as to form a carbocycle or heterocycle, W isiodo, bromo, chloro or OTf, Y is B(OH)₂ or boronate esters such asB(OCH₃)₂ and B(OC(CH₃)₂C(CH₃)₂O), iodo, SnR₃ wherein R is (C₁₋₆)alkyl,ZnX wherein X is halo, and P is a protecting group, such as commonlyused protecting groups for carboxylic acids, including, but not limitedto a methyl or ethyl ester.

Several coupling methods between the intermediate (I) (i.e. quinolinescaffold) and the intermediate II (i.e. R⁴ substituent) can becontemplated by those skilled in the art. For examples, but not limitedto, Suzuki cross-coupling between the boronic acid or boronate esterderivative of intermediate II and the halo or triflate derivative ofintermediate I, copper catalyzed Ullmann cross-coupling between the iododerivatives of intermediates I and II, Negishi cross-coupling betweenthe arylzinc reagent of the intermediate II and the iodo or triflatederivative of intermediate I, and Stille coupling between the arylltinreagent of intermediate II and the bromo or iodo derivative of I asshown above can lead, after saponification, to the compounds of formula(I).

Alternatively, the same cross-coupling methods can be used byinterchanging the coupling partners as shown below. For examples,Suzuki, Negishi, and Stille type cross-coupling between boronic acid orboronate ester derivative, the arylzinc reagent or the arylltin reagentof quinoline intermediate III and the required iodo, bromo, chloro ortriflate derivative of intermediate IV can also lead, aftersaponification, to the compounds of the invention of formula (I).

wherein R⁴², R⁴³, R⁴⁴, R⁴⁵ and, R⁴⁶ and P are as defined above and W isiodo, bromo, chloro or OTf, Y is B(OH)₂ or boronate esters such asB(OCH₃)₂ and B(OC(CH₃)₂C(CH₃)₂O), SnR₃ wherein R is (C₁₋₆)alkyl, and ZnXwherein X is halo.

Furthermore, downstream modifications to the product can becontemplated, such as conversion of an aniline-type amine to a chloro orbromo substituent via Sandmeyer reaction or alkylation, ordehalogenation via reduction.

Additionally, intermediate III can be used for decarboxylative biarylcross-coupling reactions similar to those described by Forgione,Bilodeau and coworkers, J. Am. Chem. Soc. 2006, 128, 11350-11351, hereinincorporated by reference, as shown below:

wherein W is iodo, bromo, chloro or OTf, R may be a substituent on thering and P is as defined herein.

There are a number of transformations known to access quinolinescaffolds. As shown in Scheme 1A, a Friedlander approach can be followedin which appropriately substituted aniline is condensed with afunctionalized ketone under dehydration conditions. This intermediate isthen cyclized under thermal conditions followed by halogenation of theresulting alcohol. The acetic acid ester side chain can be oxidized andprotected to furnish the alpha t-butoxy acetic acid ester moiety asshown. Separation of the enantiomers can be accomplished by formation ofdiastereomers by addition of a chiral auxiliary such as an oxazolidinonefollowed by conversion to the corresponding ester by known means.

Alternatively, a modification of this approach can also be used toprepare the quinoline scaffold as shown in Scheme 2. In this method aproperly substituted anthranilic acid derivative can be condensed underdehydration conditions with an appropriate ketone and subsequentlycyclized under DMAP/POCl₃ conditions to the 4-chloroquinoline. Furtherelaboration can then be performed as outlined in Scheme 1A.

Furthermore, in an alternative route the quinoline scaffold can beaccessed in an enantioselective manner as outlined in Scheme 3.

A quinoline precursor can be selectively brominated in the 3-positionand subsequently elaborated into the chiral diol by standard methodsknown in the literature. The chiral diol can be differentially protectedto the t-butyl ether followed by liberation of the primary alcohol. Thisalcohol can then be oxidized to the corresponding carboxylic acid andsubsequently protected as the methyl ester to furnish the key chiral4-iodoquinoline intermediate.

In an alternate route to compounds of general formula I, the knownaldehyde VIa is transformed to terminal alkyne VIb. Those skilled in theart will recognize that there are a number of methods for accomplishingthis transformation, such as, but not limited to the Bestmann-Ohirareaction or the Corey-Fuchs reaction. The R⁴ group is then attached tothe alkyne using conditions well-known to those skilled in the art,preferentially via a Sonogashira coupling between the alkyne and thearyl iodide derivative of the R⁴ group, to give the internal alkyne VIc.Other methods may include the Castro-Stevens reaction, or the silvermediated, palladium catalyzed coupling of alkyne VIb and the boronicacid or ester derivative of the R⁴ fragment as reported by Zou andcoworkers (Tetrahedron Lett. 2003, 44, 8709-8711) to give the internalalkyne VIc. In this method a properly substituted benzoylacetonitrilecan be condensed in the presence of sulfur with an appropriate ketone oraldehyde by standard methods known in the literature. VIc then undergoesa cyclocondensation with amide VId to give quinoline VIe. Those skilledin the art will recognize this may involve activation of amide VId tofacilitate the overall condensation. This is preferentially achieved bythe action of triflic anhydride and in the presence of 2-chloropyridineas described by Movassaghi (J. Am. Chem. Soc., 129 (33), 10096-10097,2007), but may also be achieved in other ways. Amides VId are typicallycommercially available, although those skilled in the art will recognizethat they are also easily obtained from commercially available anilineor nitro arene precursors. The cyclic diketal is then hydrolyzed to givediol VIf under acidic conditions. The terminal alcohol is then protectedto give VIg, where P can be a number of different protecting groupsincluding, but not limited to, a trimethylacetyl group. The secondaryalcohol is then derivatized with a tert-butyl group to give compoundVIh. Those skilled in the art will recognize that this can beaccomplished in more than one way, including an SN₁ reaction or acidcatalyzed addition to isobutylene. The protecting group is then removedto give primary alcohol VIj, which in turn is oxidized to carboxylicacid VIk. It will be obvious that the oxidation of VIj to VIk can beaccomplished in one or two synthetic steps. In the preferred method,Dess-Martin oxidation to an intermediate aldehyde followed by Lindgrenoxidation is employed.

In yet another route to compounds of general formula I, synthesis ofintermediate VIh may also be accomplished following a path that beginswith acid catalyzed hydrolysis of the cyclic diketal of terminal alkyneVIb to give diol VIIa. The terminal alcohol is then protected to giveVIIb, where P can be a number of different protecting groups including,but not limited to, a trimethylacetyl group. The secondary alcohol isthen derivatized with the tert-butyl group to give compound VIIc. Thoseskilled in the art will recognize that this can be accomplished in morethan one way, including an SN₁ reaction or acid catalyzed addition toisobutylene. The R⁴ group is then attached to the alkyne usingconditions well-known to those skilled in the art, preferentially via aSonogashira coupling between the alkyne and the aryl iodide derivativeof the R⁴ group, to give the internal alkyne VIId. The internal alkyneVIId then undergoes a cyclocondensation with amide VId to give quinolineVIh, preferentially achieved by the action of triflic anhydride and inthe presence of 2-chloropyridine as described for step 3 of Scheme 4.From intermediate VIh, the synthesis compounds of general formula I isthen accomplished following steps 7 and 8 of Scheme 4.

EXAMPLES

Other features of the present invention will become apparent from thefollowing non-limiting examples which illustrate, by way of example, theprinciples of the invention. It will be apparent to a skilled personthat the procedures exemplified below may be used, with appropriatemodifications, to prepare other compounds of the invention as describedherein.

As is well known to a person skilled in the art, reactions are performedin an inert atmosphere (including but not limited to nitrogen or argon)where necessary to protect reaction components from air or moisture.Temperatures are given in degrees Celsius (° C.). Solution percentagesand ratios express a volume to volume relationship, unless statedotherwise. Flash chromatography is carried out on silica gel (SiO₂)according to the procedure of W. C. Still et al., J. Org. Chem., (1978),43, 2923. Mass spectral analyses are recorded using electrospray massspectrometry. A number of intermediate and final products are purifiedusing CombiFlash® Companion apparatus, purchased from Teledyne Isco Inc,employing pre-packed silica gel cartridges and EtOAc and hexanes assolvents. These cartridges are available either from Silicycle Inc(SiliaFlash, 40-63 microns silica) or from Teledyne Isco (RediSep, 40-63microns silica). Preparative HPLC is carried out under standardconditions using a SunFire™ Prep C18 OBD 5 μM reverse phase column,19×50 mm and a linear gradient employing 0.1% TFA/acetonitrile and 0.1%TFA/water as solvents. Compounds are isolated as TFA salts whenapplicable. Analytical HPLC is carried out under standard conditionsusing a Combiscreen ODS-AQ C18 reverse phase column, YMC, 50×4.6 mmi.d., 5 μM, 120 Å at 220 nM, elution with a linear gradient as describedin the following table (Solvent A is 0.06% TFA in H₂O; solvent B is0.06% TFA in CH₃CN):

Time (min) Flow (mL/min) Solvent A (%) Solvent B (%) 0 3.0 95 5 0.5 3.095 5 6.0 3.0 50 50 10.5 3.5 0 100

Abbreviations or Symbols Used Herein Include:

Ac: acetyl;AcOH: acetic acid;Ac₂O: acetic anhydride;Anti-his XL665: XL665 labeled anti-His antibody;BOC or Boc: tert-butyloxycarbonyl;BSA: bovine serum albumin;Bu: butyl;CD: circular dichroismDABCO: 1,4-diazabicyclo[2.2.2]octaneDba: dibenzylidene acetone;DBU: 1,8-diazabicyclo[5.4.0]undec-7-ene;DCE: dichloroethane;DEAD: diethyl azodicarboxylate;DCM: dichloromethane;DIAD: diisopropyl azodicarboxylate;DIBAL: diisobutyl aluminum hydride;DIPEA: diisopropylethylamineDMAP: N,N-dimethyl-4-aminopyridine;DME: 1,2-dimethoxyethane;

DMF: N,N-dimethylformamide;

DMSO: dimethylsulfoxide;

Dppf: 1,1′-Bis(diphenylphosphino)ferrocene;

EC₅₀: 50% effective concentration;Eq: equivalent;Et: ethyl;Et₃N: triethylamine;Et₂O: diethyl ether;EtOAc: ethyl acetate;EtOH: ethanol;HATU: O-(7-Azabenzotriazole-1-yl)-N,N,N′N′-tetramethyluroniumhexafluorophosphate;HBTU: O-Benzotriazole-N,N,N′,N′-tetramethyluronium hexafluorophosphate;HEPES: N-2-hydroxyethyl piperazine N-ethane sulfonic acid;HPLC: high performance liquid chromatography;IC₅₀: 50% inhibitory concentration;ITC: Isothermal calorimetry;^(i)Pr or i-Pr: 1-methylethyl (iso-propyl);Kd_(app): apparent affinity constant;KHMDS: potassium hexamethyl disilazane;LiHMDS: lithium hexamethyldisilazide;Me: methyl;MeCN: acetonitrile;MeOH: methanol;MOI: multiplicity of infection;MS: mass spectrometry (ES: electrospray);n-BuONa: sodium n-butoxiden-BuOH: n-butanol;n-BuLi: n-butyl lithium;NMR: nuclear magnetic resonance spectroscopy;OD: optical density;ORD: optical rotary dispersion;Ph: phenyl;PhMe: toluene;PG: protecting group;PPh₃: triphenylphosphine;Pr: propyl;RPMI: Roswell Park Memorial Institute (cell culture medium);RT: room temperature (approximately 18° C. to 25° C.);SM: starting material;Strep-EuK: Streptavidin labeled with europium cryptate;tert-butyl or t-butyl: 1,1-dimethylethyl;TCEP: tris[2-carboxyethyl]phosphine;Tf: trifluoromethanesulfonyl;Tf₂O: trifluoromethanesulfonic anhydride;TFA: trifluoroacetic acid;THF: tetrahydrofuran; andTLC: thin layer chromatography.

Example 1 Synthesis of Quinoline Scaffold 1i

Step 1:

In a 4-neck 500 mL round bottom flask equipped with a magnetic stir bar,condenser and Dean-Stark trap, diethyl acetylsuccinate (6 g, 0.026 mol),aniline 1a (2.5 mL, 0.028 mol), Amberlyst® 15 (0.08 g) and toluene (30mL) are added. The resulting mixture is heated at reflux temperature forapproximately 3 days at which time TLC shows only traces of SM. Thereaction mixture is cooled to RT and the Amberlyst® 15 is removed byfiltration. The filtrate is concentrated in vacuo to give a suspensionof a solid in brown liquid. The filtrate is diluted with diethyl etherand cooled. The solid is filtered and the filtrate is concentrated invacuo leaving a brown oil (˜7.8 g), which contains 1b and some cyclisedintermediate. This crude intermediate is used in the next step withoutfurther purification.

Step 2:

In a 3-neck 100 mL round bottom flask a mixture of the crudeintermediate 1b (˜7.8 g) and diphenyl ether (˜50 mL) are heated quicklyin a pre-heated (250° C.) heating mantle for 6 min (internal temperaturereaches ˜250° C.) at which time the flask is removed from the heatingmantle and is stirred until the internal temperature reaches below 100°C. The reaction mixture is then mixed with hexane (15 mL), at which timea light brown solid is formed. The solid is filtered and washed withhexane (3×10 mL) to give approximately 2.4 g of the intermediatecyclised product. A portion of this sample (1.4 g, 5.87 mmol) isdissolved in phosphorus oxychloride (5 mL) and heated at reflux for 2.5h. The reaction mixture is cooled to RT and is concentrated on vacuum.The residue is treated with sodium bicarbonate powder then partitionedbetween EtOAc and water. The combined organic layer is washed withbrine, dried over anhydrous Na₂SO₄, filtered, passed through a silicagel pad and concentrated to give 1c as a crude light brown solid (˜2.35g).

Step 3:

Crude chloro quinoline 1c (1.36 g, 5.17 mmol) is dissolved in THF (20mL), and HCl in dioxane (4 M, 5.4 mL, 0.022 mol) is added to thissolution slowly. The resulting reaction mixture is stirred at RT for 40min. The solvent is then removed in vacuo and the residue is dried onvacuum. The resulting solid and NaI (3.87 g) are suspended in MeCN (20mL), and the resulting reaction mixture is heated to reflux for 16 h.The reaction mixture is cooled to RT and treated with a saturatedaqueous solution of NaHCO₃ (20 mL). The aqueous layer is extracted withDCM, and the combined organic layer is dried over anhydrous MgSO₄,filtered and concentrated to give a brown syrup. Purification by silicagel chromatography (30% EtOAc/hexanes) provides iodo quinoline 1d as anoff-white solid (1.72 g, 94% yield).

Step 4:

To a solution of KHMDS (0.5 M in toluene, 3 mL, 1.5 mmol) in THF (8 mL)at −78° C. is added a solution of 1d (0.35 g, 0.99 mmol) in THF (8 mL).As the ester is added, the solution becomes scarlet red. This is allowedto stir at −78° C. for 30 min before being treated with the Davisreagent (0.39 g, 1.5 mmol). After addition of the oxidizing agent, thesolution becomes pale yellow and this is stirred for an additional 30min at −78° C. The reaction is quenched with saturated NH₄Cl aqueoussolution (8 mL) is warmed to RT and is diluted with EtOAc. The mixtureis washed with brine and the organic phase dried (Na₂SO₄), filtered andconcentrated to afford a solid. Purification by silica gelchromatography (hexanes/EtOAc:6/4) provides 1e as a beige solid (0.50g, >98% yield).

Step 5:

To a suspension of iodoalcohol 1e (0.53 g, 1.4 mmol) in tert-butylacetate (12 mL) at RT is added perchloric acid (0.66 mL, 4.6 mmol). Thereaction is left to stir for 2 h at RT (suspension turns into a clearsolution). The reaction is quenched with water (12 mL) and basified withsolid NaHCO₃ until pH ˜6. The crude product is extracted with EtOAc(3×10 mL), washed with brine (10 mL), dried over MgSO₄, filtered andconcentrated to afford the crude product. Purification by silica gelchromatography (hexanes/EtOAc:85/15) affords 1f as a pale yellow oil(0.56 g, 91% yield).

Step 6:

Intermediate 1f (0.59 g, 1.4 mmol) is dissolved in a 2 M NaOH aqueoussolution (7 mL, 0.014 mol) with ethanol (10 mL) and is stirred for 4 hat RT. The ethanol is then removed in vacuo. The resulting residue isdiluted with water (3 mL) and acidified with 2 M HCl solution until pH˜3-4. The residue is then extracted with DCM (3×10 mL), dried overNa₂SO₄, filtered, concentrated and dried under high vacuum to afford 1gas a foamy solid (0.56 g, >98% yield).

Step 7:

To a solution of acid 1g (0.39 g, 0.97 mmol) and HBTU (0.48 g, 1.3 mmol)in anhydrous THF (5 mL) is added diisopropylethylamine (0.5 mL, 2.9mmol). The mixture is stirred for 5.5 h at 30-35° C. (internaltemperature) at which time the sodium salt of R-(+)-benzyloxazolidinone(which is prepared by adding sodium hydride (60% dispersion in mineraloil, 78 mg, 1.95 mmol) to a solution of R-(+)-benzyloxazolidinone (0.35g, 1.9 mmol) in anhydrous THF (5 mL) is added. The resulting solution isthen stirred at RT for 16 h. The solvent is removed in vacuo andpartitioned between water and EtOAc. The aqueous phase is then extractedwith EtOAc, and the combined organic extracts are dried over anhydrousNa₂SO₄, filtered, and concentrated in vacuo to afford a pale yellowsolid. The crude product is purified by silica gel chromatography(10−>30% EtOAc:hexanes), yielding the desired diastereomer 1 h (190 mg,35% yield, more polar product, >99% ee by chiral column).

Step 8:

To a solution of oxazolidinone 1 h (190 mg, 0.34 mmol) in THF/H₂O (2mL/1 mL) at 0° C. is added H₂O₂ (30%, 0.36 mL) followed by LiOHmonohydrate (17 mg, 0.41 mmol) dissolved in water (1 mL). The reactionmixture is stirred at 0° C. for 30 min at which time 10% Na₂SO₃ (0.26mL) is added. The resulting mixture is stirred for ˜10 min and thenacidified with 2 N HCl to pH ˜4-5. The product is then extracted withDCM (3×10 mL). The combined organic extracts are dried over sodiumsulfate and concentrated in vacuo to yield the crude acid intermediateas a white foam (0.13 g, 96% yield), which is used in the next stepwithout further purification. The acid (130 mg) is suspended in diethylether (3 mL) and treated with diazomethane in diethyl ether until all ofthe acid SM is consumed (as indicated by TLC). The reaction is quenchedwith a very small amount of glacial AcOH and then concentrated in vacuoto give an off-white solid. The crude ester product is purified bysilica gel chromatography (10-15% EtOAc/hexanes) yielding the quinolinefragment 1i (120 mg, 89% yield) in high enantiomeric purity (>99% ee bychiral HPLC).

Example 2 Synthesis of Fragment 2f

Step 1:

Aldehyde 2a (5.85 g, 28.6 mmol, for preparation see: Michel, P. and Ley,S. V. Synthesis 2003, 10, 1598-1602, herein incorporated by reference),phoshonate 2b (6.6 g, 34 mmol) and K₂CO₃ (8.8 g, 64 mmol) are combinedin MeOH (125 mL) and the reaction is stirred overnight at RT. Thereaction is evaporated nearly to dryness and the residue is partitionedbetween H₂O (250 mL) and EtOAc (500 mL). The water layer is washed withEtOAc (2×250 mL) and the combined organic layers dried over anhydrousNa₂SO₄ and concentrated to give alkyne 2c (5.55 g, 97% yield).

Step 2:

Alkyne 2c (5.0 g, 25 mmol) is dissolved in TFA (35 mL) and water (3.6mL) and the solution is stirred at RT. After 30 min, the reaction isconcentrated under reduced pressure and the residue is purified byCombiFlash® Companion to give diol 2d (1.8 g, 84% yield).

Step 3:

A solution of diol 2d (1.2 g, 14 mmol) and triethylamine (1.7 mL, 12mmol) in DCM (80 mL) is cooled to 0° C. under N₂.Trimethylacetylchloride is added dropwise and the resulting mixture isallowed to come to RT and stir overnight. The reaction is then quenchedwith MeOH (100 mL) and stirring is continued for 20 min. The mixture isthen concentrated under reduced pressure and the residue is purified byCombiFlash® Companion to give the desired mono ester 2e (550 mg, 40%yield) along with the undesired regioisomeric mono ester (378 mg, 27%yield).

Step 4:

In a sealable reaction flask, a solution of the propargylic alcohol 2e(375 mg, 2.20 mmol) and Amberlyst® H-15 resin (150 mg) in hexane (3 mL)is cooled to −78° C. Isobutene is then bubbled through the solutionuntil the volume approximately doubles. The tube is then sealed, broughtto RT and is stirred overnight. The tube is then cooled to −78° C., isopened and brought back to RT. The mixture is then filtered through aplug of SiO₂ (EtOAc wash) and concentrated under reduced pressure toprovide pure tert-butyl ether 2f (390 mg, 78% yield).

Example 3 Synthesis of Alkyne 3a

Step 1:

Solid Pd(PPh₃)₄ (444 mg, 0.385 mmol) and CuI (146 mg, 0.769 mmol) aresuccessively added to a solution of 6-iodochroman (10 g, 34 mmol) andalkyne 2c (11 g, 55 mmol) dissolved in DMF (23 mL) and diethylamine (115mL). The reaction mixture is stirred overnight at RT and thenconcentrated, diluted with EtOAc (300 mL) and successively washed withbrine, 1 N aqueous HCl and water (300 mL each). The organic layer isdried over Na₂SO₄ and the residue purified by CombiFlash® Companion togive alkyne 3a (10.8 g, 84% yield).

Example 4 Synthesis of Boronate Fragment 4f (Used in Preparation of1086)

Step 1:

To a solution of 4a (6 g, 37 mmol) in nitrobenzene (12 mL), chloroacetylchloride (4.6 mL, 57.5 mmol) is added, followed by the addition of AlCl₃(20.4 g, 152 mmol). As the AlCl₃ is added, the mixture becomes viscousand gas evolution is observed. The resulting brown syrupy mixture isleft to stir overnight at RT. (Reference: Y. Takeuchi et. al., Chem.Pharm. Bull. 1997, 45(12), 2011-2015.) The thick reaction mixture iscooled and ice water is added very carefully (Very exothermic!!) a fewdrops at a time. Once gas evolution and bubbling is subsided, cold wateris further added followed by EtOAc. The mixture is stirred for 5 min andthe product extracted with EtOAc (3×). The combined organic layers arewashed with brine (1×), dried over Na₂SO₄, filtered and concentrated toafford the uncyclized chloroketone (24 g of crude; contaminated withsome nitrobenzene) as a pale yellow solid. This intermediate is thentaken up in EtOH (100 mL), NaOAc is added (20.4 g, 248 mmol) and thereaction is brought to reflux for 40 min. The EtOH is evaporated, theresidue is taken up in EtOAc (300 mL) and washed with 5% K₂CO₃ (2×200mL) and the aqueous layer then acidified with aqueous HCl (1 N; pH=˜5).This acidic layer is extracted with EtOAc (2×250 mL), washed with brine(1×), dried over Na₂SO₄, filtered and concentrated to afford the crudeproduct. This material is purified by CombiFlash® Companion (120 g) toafford intermediate 4b as a yellow solid (4.7 g).

Step 2:

The ketone 4b (127 mg, 0.64 mmol) is dissolved in EtOH (2 mL) andtreated with hydrazine hydrate (500 μL, 16 mmol). The mixture is heatedto reflux for 45 min before allowing it to cool to RT. The solvent isremoved by evaporation and the residue is dissolved in diethylene glycol(1 mL) before being treated with KOH (108 mg, 1.92 mmol) and then heatedto 110-120° C. for 2.5 h. The reaction mixture is diluted with EtOAc andthe pH is adjusted with 1 N HCl to pH<4. The organic phase is separated,washed with saturated brine, dried over anhydrous MgSO₄, filtered andconcentrated. The crude material is purified by CombiFlash® Companion(eluent: 0-50% EtOAc/hexanes) to give intermediate 4c as a yellow oil(62 mg).

Step 3:

A solution of 4c (61 mg, 0.33 mmol) is cooled to −78° C. in DCM (2 mL)and then treated with BBr₃ (1 M in DCM, 825 μL, 0.82 mmol). After 15min, the bath is removed and the reaction is allowed to reach RT. Thereaction is then stirred for 1.5 h. The reaction is cooled to 0° C.before quenching by the careful dropwise addition of water. The mixtureis treated with saturated NaHCO₃ (to pH ˜8) and the phases separated.The organic phase is washed with saturated brine, dried over MgSO₄,filtered and concentrated to dryness. The product is purified byCombiFlash® Companion (0-50% EtOAc/hexanes) to give intermediate 4d ascolorless oil, which solidifies upon standing (40 mg, 71% yield).

Step 4:

The phenol 4d (40 mg, 0.23 mmol) is dissolved in DCM (2 mL), cooled to0° C. and treated with pyridine (95 μL, 1.17 mmol), followed by Tf₂O (44μL, 0.26 mmol). The reaction is allowed to stir at this temperature for10 min before warming to RT over a period of 1 h. The reaction mixtureis diluted with DCM and the organic phase washed with 10% citric acidand then brine. The organic phase is dried over anhydrous MgSO₄,filtered, concentrated and purified by CombiFlash® Companion (0-50%EtOAc/hexanes) to give 4e as a yellow oil (67 mg, 94% yield).

Step 5:

To a solution of the triflate 4e (66 mg, 0.22 mmol) in DMF (2 mL),bis-(pinacolato)diborone (72 mg, 0.28 mmol) and potassium acetate (64mg, 0.65 mmol) are added. This solution is de-gassed (with bubbling Ar)for 10 min before adding PdCl₂(dppf)-CH₂Cl₂, (27 mg, 0.03 mmol). Themixture is de-gassed a further 5 min before being heated to 90° C. for16 h. The mixture is cooled to RT and diluted with EtOAc/water. Theorganic phase is washed with saturated brine (3×), dried over anhydrousMgSO₄, filtered and concentrated. The crude material is purified byCombiFlash® Companion (0-70% EtOAc in hexanes) to afford the boronate 4fas a white solid (41 mg, 67% yield).

Example 5 Synthesis of Boronate Fragment 5f (Used in Preparation of1077, 1091, 1095, 1099, 1100, 1118)

Step 1:

The nitrophenol 5a (5.23 g, 34.1 mmol) is dissolved in acetic acid (20mL) and the solution is cooled in an ice bath. Bromine (1.75 mL, 34.15mmol), dissolved in 5 mL acetic acid) is added dropwise with stirring.The mixture is stirred for 1 h at 0° C. before being poured into icewater (250 mL). The mixture is extracted with EtOAc (2×100 mL) and thenwashed with 5% NaHCO₃ (2×50 mL) before being dried over anhydrous MgSO₄,filtered and concentrated to give the desired crude product 5b as anorange solid (8.2 g, quantitative yield). This material is used in thenext step without further purification.

Step 2:

To a well stirred ethanol solution (75 mL) of 5b (8.1 g, 34.9 mmol),SnCl₂ (20 g, 105 mmol) is added. The reaction mixture is stirred atreflux for 2.5 h. After that period, the transformation is incomplete,therefore, more SnCl₂ (2 g, 10 mmol) is added and the reaction mixtureis heated at reflux for 1 h before being cooled to RT. The mixture ispoured onto 250 g of ice and the pH adjusted to approximately 7.5 withaqueous 5% NaHCO₃. The product is extracted with EtOAc (3×100 mL) beforebeing washed with saturated brine (2×100 mL). The organic phase is driedover anhydrous MgSO₄, filtered and concentrated to dryness to give theaniline intermediate 5c as a gray solid (8.25 g, ˜100% yield; thismaterial contained some tin residues, nonetheless, it is used as suchfor the following step).

Step 3:

To a stirring, ice cold, DMF (5 mL) suspension of potassium carbonate(2.05 g, 14.8 mmol) and aniline 5c (750 mg, 3.71 mmol) under nitrogen,chloroacetyl chloride (355 μL, 4.45 mmol) is added dropwise. The mixtureis allowed to warm to RT over a period of 15 min and then heated to ˜60°C. for 1 h. The mixture is allowed to cool to RT, is poured into amixture of ice/water (250 mL) and is stirred for 15 min. The suspensionis centrifuged, and the supernatant is discarded. The solid material isleft drying under suction overnight to give intermediate 5d (280 mg, 31%yield).

Step 4:

To an ice cold THF (6 mL) solution of the cyclic amide 5d (280 mg, 1.16mmol) under nitrogen, a borane-THF solution (1M in THF, 1.74 mL, 1.74mmol) is added slowly. The reaction mixture is slowly allowed to warm toRT, then is stirred at RT for approximately 1.5 h and then gently heatedto reflux for 1 h to complete the conversion. The mixture is cooled inan ice bath and is carefully quenched with aqueous 1 M NaOH (4 mL) over10 min. The reaction mixture is partitioned between EtOAc (150 mL) andwater (25 mL). The organic layer is washed with aqueous 1 N NaOH (20mL), saturated aqueous NaCl, and finally dried over anhydrous MgSO₄,filtered and concentrated to give the crude 5e as an amber oil (212 mg,81% yield). This product is used as such for next transformation.

Step 5:

A well stirred DMF (15 mL) solution of the arylbromide 5e (0.50 g, 2.19mmol), potassium acetate (0.728 g, 7.67 mmol) andbis(pinacolato)diborane (0.83 g, 3.3 mmol) is degassed by bubbling Arthrough the solution for 20 min. PdCl₂(dppf)-DCM (320 mg, 0.44 mmol) isadded and degassing is continued for 15 min. The system is sealed(teflon screw cap vessel) under Ar and heated to ˜90° C. for 5 h. Thereaction mixture is allowed to cool to RT, dilute with EtOAc (150 mL),washed with brine (3×100 mL) and water (2×100 mL), dried over anhydrousMgSO₄, filtered and concentrated to dryness. The residue is purified byCombiFlash® Companion (EtOAc/hexanes) to give the desired boronate 5f(389 mg, 65% yield) as a yellowish waxy solid.

Example 6 Synthesis of Boronate Fragment 6i (Used in Preparation of1038, 1039, 1053, 1054, 1055, 1056, 1083)

Step 1:

Sodium hydride (60%, 7.78 g, 194 mmol) is added to a well stirredsuspension of 6a (12.5 g, 97.2 mmol) in THF (100 mL). After stirring thereaction mixture for 1 h, N,N-diethylcarbamoyl chloride (24.64 mL, 194mmol) is added at RT. After stirring the reaction overnight, thereaction mixture is quenched with water (100 mL), extracted with EtOAc(3×50 mL), is dried over anhydrous MgSO₄, filtered and evaporated underreduced pressure to obtain 6b (33 g, 75% yield) in high purity.

Step 2:

Diisopropylamine (21.0 mL, 121 mmol) in THF (330 mL) is treated with asolution of n-BuLi (2.5 M in hexanes, 48.2 mL, 121 mmol) at 0° C. After30 min at this temperature, the solution is cooled to −78° C. andcarbamate 6b (33.29 g, 109.7 mmol, 75% pure) is added. The reaction isstirred at this temperature for 30 min and then iodine (33.4 g, 132mmol) is added. The solution is stirred for 30 min at 0° C. and is thenwarmed to RT. After 2 h, the reaction mixture is quenched with water(250 mL) and the volatile organic solvents are removed under reducedpressure. The aqueous phase is then extracted with EtOAc (3×100 mL),washed with 1 N HCl (200 mL), dry MgSO₄, filtered and evaporated underreduced pressure to obtain 6c (18.6 g, 39% yield).

Step 3:

The iodocarbamate 6c (10 g, 28 mmol), propargyl alcohol (3.3 mL, 56mmol), Pd(PPh₃)₄ (3.27 g, 2.83 mmol) and copper iodide (1.08 g, 5.66mmol) are combined in diisopropylamine (39 mL, 39 mmol) in a sealabletube under Ar and heated at 100° C. After 1 h, the reaction mixture iscooled to RT and poured into EtOAc (100 mL) and this mixture isextracted with 10% HCl (2×100 mL). The organic layer is dried over MgSO₄and concentrated to dryness. The crude product is purified byCombiFlash® Companion to obtain 6d (3.65 g, 46% yield).

Step 4:

6d (3.63 g, 12.9 mmol) is dissolved in EtOAc (81 mL) and treated withRh—Al₂O₃ (5% w/w, 3.45 g, 1.68 mmol). The flask is evacuated and chargedwith 1 atmosphere of H₂ (balloon) and the reaction is stirred overnightat RT. The reaction mixture is filtered through Celite® (EtOAc wash) andthe filtrate is concentrated under reduced pressure. The residue is thenpurified by CombiFlash® Companion to obtain 6e (3.7 g, 71% yield).

Step 5:

Solid NaOH (920 mg, 23 mmol) is added to a solution of 6e (2.63 g, 9.20mmoL) in EtOH (93 mL) and the mixture is heated to reflux and is stirredovernight. The mixture is then cooled to RT and the organic solventremoved under reduced pressure. Water is added (100 mL) and the mixtureextracted with Et₂O (3×100 mL), dried over MgSO₄, filtered andevaporated under reduced pressure to obtain phenol 6f (869 mg, 51%yield).

Step 6:

Diethyl azodicarboxylate (953 μL, 6.05 mmol) is added dropwise to asolution of phenol 6f (869 mg, 4.66 mmol) and PPh₃ (1.59 g, 6.053 mmol)in THF (65 mL) and the reaction is stirred at RT. After 4 h, thereaction mixture is evaporated under reduced pressure. The residue isthen purified by CombiFlash® Companion to obtain the chroman 6g (387 mg,49% yield).

Step 7:

Iodine (583 mg, 2.295 mmol) is added to a solution of chroman 6g (387mg, 2.29 mmol) and AgNO₃ (429 mg, 2.52 mmol) in MeOH (23 mL). After 20min, a 0.5 M solution of sodium thiosulfate (10 mL) is added and theaqueous phase extracted with EtOAc (3×25 mL). The combined organicphases are washed with brine, then dried (MgSO₄), filtered andevaporated to obtain aryl iodide 6h (647 mg, 96% yield).

Step 8:

A solution of iodo intermediate 6h (647 mg, 2.20 mmol),bis(pinocolato)diborane (0.725 g, 2.86 mmol) and potassium acetate(0.626 g, 6.59 mmol) in DMF (17 mL) is degassed with Ar for 10 min.PdCl₂(dppf)-DCM complex (179 mg, 0.22 mmol) is then added and themixture is degassed with Ar for approximately another 5 min. Thereaction is then heated to 95° C. in a sealable tube and is stirredovernight. The reaction is cooled to RT and EtOAc (100 mL) is added. Thesolution is washed with brine (3×150 mL), water (1×150 mL), dried overMgSO₄, filtered and solvent removed under reduced pressure. The residueis purified by CombiFlash® Companion to afford boronate ester 6i (260mg, 40% yield).

Example 7 Synthesis of Boronate Fragment 7d (Used in Preparation of1065, 1107)

Step 1:

A solution of phenol 7a (0.91 g, 5.74 mmol) in dry DMF (1 mL) is addeddropwise to a slurry of NaH (60% in oil, 0.60 g, 15 mmol) in dry DMF (1mL) cooled to 10-15° C. (cold water bath) and the mixture is stirred for20 min. This results in a thick, frothy white mixture. A solution of3-bromopropionic acid (1.1 g, 6.9 mmol) in dry DMF (0.5 mL) is thenadded dropwise and the reaction stirred at RT overnight. After 16 h,MeOH (1.2 mL) is added to help break up the thick, pasty reactionmixture which is then added to diluted HCl (˜12 mL, 1 N HCl in 100 mLwater) and extracted with EtOAc (80 mL; the pH of the aqueous phase isadjusted to pH<3). The organic layer is dried over anhydrous Na₂SO₄ andevaporated to give 7b as a white solid material, contaminated with someunreacted SM (1.29 g of crude material). This material is used in thenext step without purification.

Step 2:

The crude compound 7b (1.53 g, 6.63 mmol) is combined withpolyphosphoric acid (approximately 7 g) and heated to 75° C. to give acherry red colored solution. During the reaction time, the reactionmixture becomes viscous and stirring becomes difficult. After 4 h, iceand water are slowly added with rapid stirring to give a thicksuspension. This mixture is transferred to a separatory funnel where theproduct is extracted with EtOAc (100 mL) and washed with water (100 mL),saturated NaHCO₃ (2×100 mL) and brine (75 mL). The organic phase isdried over anhydrous MgSO₄ and evaporated to give 7c as a sticky violetsolid which is used as such (1.29 g).

Step 3:

Intermediate 7c is analogous to intermediate 4b in Example 4; thoseskilled in the art would recognize that the same synthetic methodologiesused to convert 4b to the boronate 4f can be applied for the conversionof 7c to the corresponding boronate 7d.

Example 8 Synthesis of Boronate Fragment 8h (Used in Preparation of1069)

Step 1

2-Amino-m-cresol 8a (5.7 g, 46.3 mmol) is dissolved in H₂O (30 mL) and1,4-dioxan (15 mL). The mixture is heated to reflux and then HBr (48%,17 mL, 0.31 mol) is added dropwise over a period of 20 min. The refluxis maintained for an additional 15 min after the addition is complete.The reaction is cooled to 0° C., and NaNO₂ in H₂O (20 mL) is added overa period of 30 min. The stirring is continued for 15 min at 0° C., themixture is then transferred in one shot to a stirring mixture of Cu(I)Br(7.64 g, 53.2 mmol) in H₂O (20 mL) and HBr (48%, 17 mL, 0.31 mol) at 0°C. (protected from light). The reaction is stirred for 15 min at 0° C.,warmed to 60° C., stirred for an additional 15 min, cooled to RT andthen stirred overnight. The reaction mixture is then transferred to aseparatory funnel and extracted with EtOAc (3×). The organic layers arecombined, washed with brine, dried over anhydrous MgSO₄, filtered andconcentrated over silica to afford a mixture that is purified using theCombiFlash® Companion (20% EtOAc/hexanes) to afford the desired bromide8b (1.46 g, 17% yield) as a red-brown oil.

Step 2:

To a solution of the bromide 8b (1.36 g; 7.27 mmol) and (PPh₃)₂PdCl₂(766 mg, 1.09 mmol, 15 mol %) in DMF (12 mL),1-ethoxyvinyl-tri-n-butyltin (2.7 mL, 8.0 mmol) is added. The mixture iscapped and heated in a microwave at 160° C. for 15 min. HPLC and LC-MSanalysis indicate approximately 70% conversion. More1-ethoxyvinyl-tri-n-butyltin (2.7 mL; 8.0 mmol) and catalyst(PPh₃)₂PdCl₂ (380 mg, 0.05 mol %) are added and the solution is againsubjected to the same microwave conditions. The reaction is quenchedwith 6N HCl (1.5 mL) and stirred at RT for 1 h to effect hydrolysis ofthe intermediate. The mixture is poured into EtOAc (150 mL), washed withbrine (3×), dried over MgSO₄, filtered and concentrated over silica toafford the mixture that is purified using the CombiFlash® Companion toafford the desired ketone 8c (947 mg, 87% yield) as an orange oil.

Step 3:

The methyl ketone 8c (1.02 g, 6.8 mmol) is dissolved in EtOAc (15 mL)and CHCl₃ (15 mL) before being treated with Cu(II)Br₂ (3.03 g, 13.6mmol). The mixture is heated to reflux for 16 h. The mixture is cooledto RT, the product filtered and washed with EtOAc (1×). The solution isconcentrated over silica to afford the mixture that is purified usingthe CombiFlash® Companion (10% EtOAc/hexanes) to afford theα-bromoketone 8d (710 mg, 46% yield) as an orange oil.

Step 4:

To a solution of the bromoketone 8d (710 mg, 3.1 mmol) in anhydrous DMF(12 mL), KF (400 mg, 6.95 mmol) is added. The reaction is stirred at RTfor 16 h. The mixture is taken up in EtOAc (150 mL), washed with brine(3×), dried over anhydrous MgSO₄, filtered and concentrated over silicato afford the mixture that is purified using the CombiFlash® Companion(20% EtOAc/hexanes) to afford the cyclic ketone 8e (280 mg, 61% yield)as a pale orange solid.

Step 5:

Zn dust pre-activation procedure: Zinc dust (20 g, 350 mesh) is placedin a round bottom flask and 1 N HCl (50 mL) is added. This suspension issonicated for 1 min before decanting off the liquid. This procedure isrepeated for a second time after which the solid is washed with EtOH(2×), Et₂O (2×) and dried under high vacuum. To a solution of the ketone8e (280 mg, 1.89 mmol) in AcOH (10 mL) pre-activated Zn dust (1.24 g,18.9 mmol) is added. The reaction mixture is then heated to 75° C. for 2h. The reaction mixture is filtered (with EtOAc washing of the solids).The solvent is evaporated over silica and the mixture is directlypurified using the CombiFlash® Companion (10% EtOAc/hexanes) to affordthe desired dihyrobenzofuran 8f (174 mg, 69% yield) as a colorless oil.

Step 6:

To a solution of the dihydrobenzofuran 8f (240 mg, 1.8 mmol) in MeOH (5mL), AgNO₃ (304 mg, 1.79 mmol) is added followed by iodine (453 mg, 1.79mmol). The yellow mixture is stirred at RT for 1 h. To the reactionmixture is added a solution of 10% Na₂S₂O₃ and the mixture is stirredfor 15 min at RT. The mixture is diluted with EtOAc (100 mL), and theorganic layer is washed with brine (3×) and 10% Na₂S₂O₃ (2×). Theorganic phase is dried over anhydrous MgSO₄, filtered and concentratedover silica to give a mixture. This mixture is purified using theCombiFlash® Companion (10% EtOAc/hexanes) to afford the iodo derivative8g (400 mg, 86% yield) as a white amorphous solid.

Step 7:

A mixture of the iodo derivative 8g (400 mg, 1.54 mmol),bis(pinocolato)diborane (585 mg, 2.31 mmol), potassium acetate (511 mg,5.4 mmol) in DMF (20 mL) is deoxygenated (Ar balloon and sonication for5 min); then the catalyst (PdCl₂dppf, 188 mg, 0.23 mmol) is added withadditional degassing (Ar balloon and sonication for 2 min). The mixtureis then heated to 95° C. for 4 h. The mixture is cooled, EtOAc (200 mL)is added, washed with brine (3×), water (2×), dried over anhydrousMgSO₄, filtered and solvent evaporation over silica affords the mixturethat is purified using the CombiFlash® Companion (10% EtOAc/hexanes) toafford the desired boronate 8h (315 mg, 79% yield) as a yellow oil.

Example 9 Synthesis of Boronate Fragment 9b (Used in Example 43)

Anhydrous DMF (60 mL) is added to a flask charged with bromide 9a (5.00g, 22.2 mmol), bis-(pinacolato)diborone (8.48 g, 33.4 mmol) andpotassium acetate (6.35 g, 66.8 mmol) and the resulting suspension isdeoxygenated by bubbling a stream of N₂ gas through the mixture for 45min. 1,1′-bis(diphenylphosphino)ferrocene (2.73 g, 3.34 mmol) is thenadded and the mixture is deoxygenated for approximately a further 5 minand is then heated to 95° C. After 16 h, the dark reaction mixture iscooled, extracted with EtOAc (500 mL and 300 mL) and washed with 1:1water/brine (600 mL) and brine (600 mL). The combined extracts are driedover anhydrous MgSO₄, filtered and evaporated to a black syrup which ispurified by flash column chromatography (EtOAc/hexanes) to afford theboronate 9b as white solid contaminated with <25% of the diboron reagent(4.24 g, 62% yield).

Example 10 Synthesis of Boronate Fragment 10g (Used in Preparation of1102, 1108, 1109, 1110, 1111, 1119, 1142)

Step 1:

2-Chloro-6-fluoronitrobenzene 10a (6.62 g, 37.7 mmol) and LiOHmonohydrate (6.33 g, 151 mmol) are dissolved in THF (45 mL) and water(65 mL) and an aqueous solution of H₂O₂ (30%, 8.60 mL, 80.0 mmol) added.The resulting turbid solution is sealed and is heated to 60° C. withrapid stirring. After 3 days, the dark orange mixture is cooled and isadded to half-saturated aqueous sodium thiosulfate (200 mL) and shakenvigorously in a separatory funnel. The mixture is then acidified to pH<3with 1 N HCl, extracted with EtOAc (400 mL+100 mL) and washed with brine(400 mL). The combined extracts are dried over magnesium sulfate,filtered and evaporated to a deep yellow oil (aminophenol 10b)containing some solid particles (residual starting material) which isused as such (6.37 g, 97% yield).

Step 2:

The crude aminophenol 10b (6.37 g, 36.7 mmol) is dissolved in THF (100mL) and tin powder (17.4 g, 147 mmol) is added followed by 1 N HCl (220mL, 220 mmol). The resulting mixture is stirred vigorously at RT. After16 h, the reaction is cooled to 0° C., the acid neutralized with 10 NNaOH (22 mL) and the resulting milky suspension stirred vigorously for15 min. The mixture is then filtered through a pad of Celite® and thesolids washed thoroughly with EtOAc (4×200 mL). The filtrate istransferred to a separatory funnel and the aqueous phase acidified with1 N HCl (4 mL), diluted with brine (400 mL) and the organic phase washedwith brine (400 mL). The extract is then dried over sodium sulfate,filtered and evaporated to afford aminophenol 10c as a waxy, pale brownsolid (2.91 g, 55% yield).

Step 3:

Chloroacetyl chloride (1.94 mL, 24.3 mmol) is added to an ice-coldmixture of aminophenol 10c (2.91 g, 20.3 mmol) and potassium carbonate(8.40 g, 60.8 mmol) in anhydrous DMF (200 mL) under a N₂ atmosphere.After 5 min, the reaction is allowed to warm to RT and, after a further45 min, is heated to 50° C. After 15 h, the reaction is cooled andextracted with EtOAc (600 mL) and washed with water/brine (1 L),half-saturated sodium bicarbonate (1 L) and brine (600 mL). The organicphase is then dried over MgSO₄, filtered and evaporated to afford lactam10d as a fibrous, pale-olive solid (3.15 g, 85% yield).

Step 4:

Bromine (1.8 mL; 35 mmol) is slowly added dropwise to a stirred solutionof lactam 10d (3.15 g; 17.1 mmol) in anhydrous DCM (40 mL) at RT. After3 h, the resulting suspension is slowly added to saturated aqueoussodium thiosulfate (200 mL) and extracted with DCM (4×100 mL). Thecombined extracts are then washed with brine (200 mL), dried overmagnesium sulfate, filtered and evaporated to afford the bromide 10e asa pale beige powder (4.00 g, 89% yield).

Step 5:

A solution of borane in THF (1.0 M, 18.5 mL, 18.5 mmol) is addeddropwise to an ice-cold solution of lactam 10e (4.00 g, 15.2 mmol) inanhydrous THF (75 mL), and the reaction is allowed to warm to RT. After30 min, the solution is heated to gentle reflux under a N₂ atmosphere.After 2 h, the reaction is cooled to 0° C. and carefully quenched with 1N NaOH (19 mL) and stirred for 15 min. The mixture is then diluted withwater (30 mL) and the THF is evaporated. The aqueous residue is thenextracted with EtOAc (400 mL+50 mL) and washed with water/brine (200mL), 0.5 N NaOH (200 mL) and brine (100 mL). The combined extracts aredried over magnesium sulfate, filtered and evaporated to afford themorpholine derivative 10f as a yellow syrup (3.90 g, quantitative.yield).

Step 6:

Anhydrous DMF (30 mL) is added to a flask charged with aryl bromide 10f(1.84 g, 7.42 mmol), bis(pinacolato)diborane (2.83 g, 11.1 mmol) andpotassium acetate (2.47 g, 26.0 mmol) and the resulting suspension isthen deoxygenated by bubbling a stream of N₂ gas through the mixture for15 min. 1,1′-bis(diphenylphosphino)ferrocene (909 mg, 1.11 mmol) is thenadded and the mixture is deoxygenated for a further 5 min and thenheated to 95° C. After 16 h, the dark reaction mixture is cooled,diluted with EtOAc (300 mL) and washed with 1:1 water/brine (500 mL) andbrine (200 mL). The extract is then dried over MgSO₄, filtered andevaporated to a brown syrup which is chromatographed over silica gel(EtOAc/hexanes) to afford the boronate 10g as a white solid contaminatedwith 0.8 eq of the diboron reagent (1.52 g, 69% yield).

Example 11 Synthesis of Boronate Fragment 11d (Used in Preparation of1006, 1021, 1022)

Step 1:

Commercially available chromanone 11a (9.78 g, 66.0 mmol) dissolved inAcOH (20 mL) is added to a suspension of zinc dust (108 g, 1.65 mol) inAcOH (150 mL). The mixture is heated to 100° C. and is stirredmechanically overnight. The mixture is then filtered through Celite®(washed with EtOAc, 100 mL), diluted with PhMe (300 mL) and the solutionis evaporated to give chroman intermediate 11b (8.45 g, 95% yield).

Step 2:

AgNO₃ (12.0 g, 70.6 mmol) and I₂ (15.8 g, 62.3 mmol) are addedsequentially to a solution of 11b (8.45 g, 63.0 mmol) dissolved in MeOH(225 mL). The reaction is allowed to stir for 1 h, filtered on Celite®and the filtrate concentrated under reduced pressure. The crude mixtureis diluted with EtOAc (250 mL) and washed with saturated sodiumthiosulfate (250 mL). The organic layer is washed with water (200 mL)and then dried over Na₂SO₄, filtered and concentrated. The crude mixtureis further purified by CombiFlash® Companion to give 6-iodochroman 11e(12.1 g, 74% yield).

Step 3:

A solution of the 6-iodochroman 11e (1.0 g, 3.85 mmol),bis[pinocolato]diborane (1.22 g, 4.81 mmol) and potassium acetate (1.10g, 11.5 mmol) in DMF (36 mL) is degassed with Ar for 5 min followed bythe addition of the PdCl₂dppf-DCM complex (314 mg, 0.38 mmol). Thereaction mixture is then degassed for an additional 5 min before beingheated to 95° C. for 5 h. The reaction is then cooled to RT. The crudereaction mixture is diluted with water and the product is extracted 3times with EtOAc (3×100 mL). The combined organics are washed with water(100 mL) and brine (100 mL). The organic phase is then dried over MgSO₄and filtered and concentrated. The crude mixture is further purified byCombiFlash® Companion using a gradient of EtOAc/hexanes to afford theborane fragment 11d (840 mg, 84% yield).

Example 12 Synthesis of Boronate Fragment 12g (Used in Preparation of1037, 1042)

Step 1:

The phenol 12a (6.75 g, 47.3 mmol) is dissolved in DMF (270 mL) and istreated with allyl bromide (6.55 mL, 75.7 mmol). To this solution, NaH(60%, 4 g, 99.4 mmol) is added portionwise and stirring is continuedovernight. The reaction mixture is diluted with EtOAc (500 mL) andwashed with H₂O (3×500 mL). The organic layer is dried over MgSO₄,filtered and concentrated to dryness to obtain the desired product 12b,which is used as such in the next step.

Step 2:

The ether 12b (9.67 g) is placed in a microwave vial neat with a stirbar and is heated to 240° C. for 20 min at which point the Claisenrearrangement reaction is complete. The crude product 12c (9.3 g) isused in the following step without further purification.

Step 3:

To a solution of the allyl intermediate 12c (9.3 g, 45.8 mmol) inanhydrous THF (300 mL) at 0° C., borane (1 M in THF, 96 mL, 96 mmol) isadded. The solution is allowed to warm to RT and then is stirred for 2.5h. The solution is then cooled to 0° C. and treated with 10 N NaOHdropwise, followed by slow addition of 30% H₂O₂ (104 ml, 916 mmol, 20eq). The resulting mixture is allowed to warm to RT and then is stirredat RT for 1 h. The reaction mixture is diluted with HCl (10%, 100 mL)and extracted with EtOAc (3×200 mL). The combined organic phases aredried over MgSO₄ and concentrated. The crude product is purified byCombiFlash® Companion to give 12d (7.1 g, 77% yield).

Step 4:

To a solution of the diol 12d (7.1 g, 35.3 mmol) in THF (500 mL), PPh₃(12 g, 45.9 mmol), followed by DEAD (7.2 mL, 45.9 mmol) are added. Thesolution is stirred at RT for 4 h. The reaction mixture is evaporatedunder reduced pressure and purified by CombiFlash® Companion to obtainthe desired product 12e (5.26 g, 82% yield).

Step 5:

The chroman derivative 12e (5.26 g, 28.8 mmol) is dissolved in AcOH (70mL) and is then treated with Br₂ in AcOH (40 mL). The reaction isstirred at RT for 15 min, then diluted with toluene and concentrated todryness. The residue is taken up in EtOAc (25 mL) and washed withsaturated Na₂S₂O₃ (25 mL) and saturated NaHCO₃ (25 mL). The organiclayer is dried over MgSO₄, concentrated and purified by CombiFlash®Companion to obtain the desired product 12f (2.7 g, 36% yield).

Step 6:

The bromide 12f (2.71 g, 10.4 mmol) is dissolved in DMF (120 mL) andtreated with bispinocolatoborane (4 g, 15.5 mmol) and potassium acetate(3.45 g, 36.3 mmol). The mixture is degassed (using an Ar balloon)before the introduction of the catalyst (PdCl₂dppf, 845 mg, 1.04 mmol).The mixture is then degassed again (using an Ar balloon) and heated to95° C. for 16 h. The mixture is cooled to RT, diluted with H₂O (300 mL)and extracted with EtOAc (2×300 mL). The combined organic layers arewashed with water (3×300 mL) dried over MgSO₄, filtered andconcentrated. The product is then purified by CombiFlash® Companion. Thesemi-purified product is then triturated with hexanes (3×50 mL) in orderto remove the excess disborane and obtain clean compound 12g (1.74 g,54% yield).

Example 13 Synthesis of Boronate Fragment 13a (Used in Preparation of1043, 1044, 1090, 1092, 1096, 1122)

Step 1:

Palladium on activated charcoal (10% Pd by weight, 0.63 mg, 0.59 mmol)is added to a solution of aryl chloride 12g (0.91 g, 2.95 mmol) andammonium formate (1.92 g, 30.4 mmol) dissolved in MeOH and the mixtureis heated to reflux. After 15 min, the reaction is cooled to RT andfiltered through Celite® (MeOH rinse). The filtrate is evaporated todryness and the residue partitioned between water and EtOAc (10 mLeach). The organic layer is dried over anhydrous MgSO₄ and concentratedto obtain boronic ester 13a (0.78 g, 97% yield).

Example 14 Synthesis of Boronate Fragment 14g (Used in Preparation of1017)

Step 1:

Allyl bromide (9.3 mL, 110 mmol) followed by potassium carbonate (20 g,150 mmol) are added to a solution of 14a (10 g, 73 mmol) dissolved inDMF (110 mL). The reaction is allowed to stir under Ar at RT overnight.The reaction is diluted with water (400 mL) and extracted with EtOAc(400 mL). The organic layer is washed with water (2×400 mL), dried overNa₂SO₄ and concentrated. The product is then purified by CombiFlash®Companion in two batches (120 g column) to provide allyl ether 14b (12g, 92% yield).

Step 2:

A solution of n-BuLi in hexanes (2.5 M, 6.4 mL, 16 mmol) is addeddropwise to a precooled (−78° C.) suspension ofmethyltriphenylphosphonium bromide (6.6 g, 19 mmol) in THF (90 mL). Theresulting bright yellow mixture is stirred for 5 min at −78° C., warmedto RT over approximately 5 min and then recooled to −78° C. Aldehyde 14b(2.4 g, 14 mmol) dissolved in THF (10 mL) is added dropwise and thereaction is allowed to proceed for 10 min at −78° C. before beingallowed to warm to RT and stir overnight. The reaction is quenched withbrine (100 mL), diluted with water (100 mL) and extracted with EtOAc(100 mL). The organic layer is then washed with water (2×100 mL), driedover Na₂SO₄ and concentrated. The crude yellow liquid is then taken upin EtOAc (1 mL) and diluted with hexanes (20 mL), after which Ph₃POprecipitates as a white solid. The solid is removed by filtration,washed with 1:9 EtOAc/hexanes (50 mL) and the filtrates are evaporatedto dryness. The product is purified by CombiFlash® Companion to givediene 14c (1.3 g, 54% yield).

Step 3:

Grubb's second generation catalyst (50 mg, 0.075 mmol) is added to adegassed solution of diene 14c (1.3 g, 7.5 mmol). After stirring underAr for 2.5 h, the reaction is concentrated onto SiO₂ (2 g) and theproduct purified by CombiFlash® Companion to give benzopyran 14d (940mg, 86% yield) as a clear oil.

Step 4:

Solid Pd—C (10% w/w, 680 mg, 0.64 mmol) is added to a solution ofbenzopyran 14d (940 mg, 6.4 mmol) in EtOH (8.5 mL) and the flask isevacuated and backfilled with H₂ gas (balloon). After stirring thereaction at RT for 2.5 h, the mixture is filtered through Celite® (EtOAcwashing) and then the filtrate is concentrated to dryness. The productis purified by CombiFlash® Companion to provide chroman 14e (800 mg, 84%yield).

Step 5:

Neat Br₂ (275 μL, 5.4 mmol) is added dropwise to a solution of chroman14e (800 mg, 5.4 mmol) dissolved in AcOH (25 mL). The reaction is thendiluted with water (50 mL) and EtOAc (50 mL). The organic layer iswashed with water (2×50 mL) and saturated NaHCO₃ (2×50 mL). The organiclayer is dried over Na₂SO₄ and concentrated to dryness. The product ispurified by CombiFlash® Companion to give bromide 14f as a mixture withthe dibromide (1.3 g, 68% by mass 14f, 51% yield).

Step 6:

A solution of the bromide 14f (950 mg, 2.8 mmol),bis[pinocolato]diborane (840 mg, 3.3 mmol) and potassium acetate (920 g,9.6 mmol) in DMF (30 mL) is degassed with Ar for 5 min followed by theaddition of the PdCl₂dppf-DCM complex (290 mg, 0.36 mmol). The reactionmixture is then degassed for an additional 5 min before being heated to95° C. for 3 h. The reaction is then cooled to RT. The crude reactionmixture is diluted with water and the product is extracted with EtOAc(3×20 mL). The combined organics are washed with water (2×20 mL). Theorganic phase is then dried over Na₂SO₄, filtered and concentrated. Thecrude mixture is further purified by CombiFlash® Companion to affordboronic ester 14g (403 mg, 53% yield) as a pale yellow solid.

Example 15 Synthesis of Boronate Fragment 15l (Used in Preparation of1098)

Step 1:

An ethereal solution of diazomethane (0.7 M, 100 mL) is added to asolution of 15a (5.0 g, 30 mmol) in ether (20 mL). After consumption ofthe SM (TLC monitoring), the reaction is concentrated onto SiO₂ (10 g)and the product purified by CombiFlash® Companion to yield ester 15b(5.2 g, 95% yield).

Step 2:

A solution of NaNO₂ (2.1 g, 30 mmol) in water (10 mL) is slowly added toa solution of aniline 15b (5.0 g, 28 mmol) dissolved in AcOH (50 mL) and2 M HCl (75 mL) at 0° C. The resulting mixture is stirred at thistemperature for 1 h. Solid CuCl (8.4 g, 85 mmol) is added portionwise(over 2 min). The reaction is allowed to come to RT, is stirred for 30min and then is warmed to 60° C. for 40 min. The mixture is poured intowater (200 mL) and extracted with EtOAc (2×200 mL). The organic layer isdried with MgSO₄, filtered and evaporated to dryness. The product ispurified by CombiFlash® Companion to afford aryl chloride 15c (3.8 g,68% yield).

Step 3:

A solution of DIBAL in DCM (1 M, 42 mL, 42 mmol) is added dropwise overa period of 25 min to a precooled (−78° C.) solution of ester 15c (3.8g, 19 mmol) in dry CH₂Cl₂ (100 mL). The reaction is allowed to stir for2 h at −78° C. The reaction is quenched at −78° C. by the dropwiseaddition of 1 N HCl (8 mL). The reaction is allowed to warm to RT andthe organic phase washed with a 5% solution of Rochelle's salt (100 mL),dried over MgSO₄, filtered and concentrated under reduced pressure togive crude benzyl alcohol 15d (3.2 g, 99% yield), which is used in thenext step without any further purification.

Step 4:

Solid Dess Martin reagent (8.7 g, 20 mmol) is added to a precooled (0°C.) solution of alcohol 15d in dry CH₂Cl₂ (100 mL). The reaction isallowed to stir for 2 h while slowly warming to RT. At this time,another 0.5 g of Dess Martin Periodinane is added and the reactioncontinues for another 1 h. A 1:1 mixture of saturated NaHCO₃ and 0.5 MNa₂S₂O₃ (100 mL) is added and this mixture is stirred vigorously untilthe phases become clear (approximately 30 min). The organic phase isseparated and the aqueous phase is extracted with DCM (100 mL) andwashed with saturated NaHCO₃ (100 mL). The combined organic phases arethen dried over MgSO₄ and evaporated. The product is purified byCombiFlash® Companion to give aldehyde 15e (2.9 g, 90% yield).

Step 5:

A solution of methyl ether 15e (720 mg, 4.2 mmol) in anhydrous CH₂Cl₂(20 mL) is added slowly to a precooled (−30° C.) solution of BBr₃ (1 M,8.4 mL, 8.4 mmol). The solution is warmed to 0° C. and is stirred for 3h. The reaction is quenched carefully with methanol (1 mL) and washedwith saturated NaHCO₃ and then brine (25 mL each). The organic layer isdried over MgSO₄, filtered and concentrated and the product is purifiedby CombiFlash® Companion to give phenol 15f (530 mg, 80% yield).

Step 6:

A mixture of the aldehyde 15f (1.1 g, 7.2 mmol), acrylonitrile (2.4 mL,36 mmol) and DABCO (190 mg, 1.7 mmol) are refluxed for 5 h. The reactionmixture is cooled to RT, diluted with EtOAc (50 mL) and washed with 1 NNaOH (20 mL) and then with 1 N HCl (20 mL). The organic phase is driedover MgSO₄ and concentrated to dryness. The product is purified byCombiFlash® Companion to afford the nitrile 15g (650 mg, 47% yield).

Step 7:

A mixture of nitrile 15g (650 mg, 3.4 mmol), 10% NaOH (10 mL, 25 mmol)and EtOH (95%, 0.5 mL) is heated to reflux for 5 days. The reaction isthen cooled to RT and 1 N HCl is then added until pH ˜4. The precipitateis then collected by filtration, washed with water and dried in vacuo togive acid 15h (740 mg, >99% yield).

Step 8:

Triethylamine (0.56 mL, 4.0 mmol) and diphenylphosphoryl azide (0.75 mL,3.5 mmol) are added successively to a solution of acid 15h (714 mg, 3.4mmol) in dry toluene (40 mL). This mixture is heated to 85° C. for 2 hand then cooled to RT and treated with 6 N HCl (6 mL). The mixture isbrought to reflux and is stirred at this temperature for 2 h. Thereaction is then cooled to RT, diluted with EtOAc (100 mL) and washedwith saturated NaHCO₃ (2×100 mL), water (2×100 mL) and brine (100 mL).The organic layer is dried over MgSO₄, filtered and evaporated todryness. The product is then purified by CombiFlash® Companion to giveketone 15i (269 mg, 44% yield).

Step 9:

Deoxofluor® (0.54 mL, 2.9 mmol) is added to a solution of ketone 15i(270 mg, 1.5 mmol) in CH₂Cl₂ (0.6 mL) and EtOH (17 μL) in a sealed tube.The sealed tube is heated to 40° C. for 24 h. The tube is then unsealed,cooled to 0° C. and the reaction quenched by the slow (Caution!Exothermic!) addition of saturated NaHCO₃ (1 mL). The crude reactionmixture is diluted with water (20 mL) and extracted with DCM (3×20 mL).The combined organics are washed with water (20 mL) and the organicphase is dried over MgSO₄, filtered and concentrated. The product ispurified by CombiFlash® Companion to provide difluorochroman 15j (225mg, 71% yield).

Step 10:

Solid silver nitrate (187 mg, 1.1 mmol) and iodine (279 mg, 1.1 mmol)are added successively to a solution of difluorochroman 15j (225 mg, 1.1mmol) dissolved in MeOH (7.8 mL). The reaction is stirred at RT for 90min and then filtered through a pad of Celite®. The filtrate is treatedwith a drop of 0.5 N Na₂S₂O₃ (orange color dissipated) then concentratedunder reduced pressure. The residue is partitioned between H₂O, 0.5NNa₂S₂O₃ and EtOAc (20 mL each). The water layer is extracted with EtOAc(3×20 mL) and the combined organics are washed with brine (20 mL), driedover MgSO₄, filtered and concentrated. The product is purified byCombiFlash® Companion to give aryl iodide 15k (158 mg, 44% yield).

Step 11:

A solution of the aryl iodide 15k (150 mg, 0.45 mmol),bis[pinocolato]diborane (150 mg, 0.59 mmol) and potassium acetate (130mg, 1.4 mmol) in DMF (5 mL) is degassed with Ar for 5 min followed bythe addition of the PdCl₂dppf-DCM complex (44 mg, 0.054 mmol). Thereaction mixture is then degassed for an additional 5 min before beingheated to 85° C. for approximately 9 h. The reaction is then cooled toRT. The crude reaction mixture is diluted with water and the product isextracted with EtOAc (3×10 mL). The combined organics are washed withwater (10 mL) and brine (10 mL). The organic phase is then dried overMgSO₄ and filtered and concentrated. The crude mixture is furtherpurified by CombiFlash® Companion to afford boronic ester 15l (123 mg,70% pure by NMR, 57% yield).

Example 16 Synthesis of Boronate Fragment 16c (Used in Preparation of1088)

Step 1:

Solid NaBH₄ (342 mg, 9.0 mmol) is added to a solution of ketone 4b (1.5g, 7.5 mmol) dissolved in MeOH (10 mL) and THF (25 mL) at 0° C. is thenadded. The reaction is warmed to RT and is allowed to stir for 1 h. Thereaction is quenched with aqueous HCl (1 N, 5 mL), the MeOH is removedby concentration and the product extracted with EtOAc (2×50 mL). Theorganic layer is washed with brine (50 mL), dried over Na₂SO₄, filteredand concentrated to afford alcohol 16a (1.52 g>99% yield). This materialis used as is in the next step.

Step 2:

TFA (2.9 mL) is added dropwise to a solution of crude alcohol 16a (1.5g; 7.47 mmol) in CH₂Cl₂ (28 mL) at 0° C. The solution is stirred for 30min, and then concentrated to dryness. The residue is taken up in EtOAc,washed with NaHCO₃ (saturated), brine, dried over Na₂SO₄, filtered andconcentrated to a pale yellow gum. The product is purified byCombiFlash® Companion to afford benzofuran 16b (0.30 g, 22% yield) as awhite solid.

Step 3:

Compound 16c is prepared from 16b following a synthetic sequenceidentical to steps 3 to 5 of Example 4.

Example 17 Synthesis of Boronate Fragment 17g (Used in Preparation of1047, 1048, 1049)

Step 1:

Zn dust (7.89 g, 121 mmol) is added to a solution of 17a (5.0 g, 24mmol) in AcOH (100 mL). The reaction mixture is then heated to 100° C.and is stirred overnight. The reaction is cooled to RT and the mixtureis filtered (EtOAc washing), the solvent is evaporated and the residuepurified by CombiFlash® Companion to afford aniline 17b (3.06 g, 72%yield) as a yellow solid.

Step 2:

A solution of NaNO₂ (640 mg, 9.3 mmol) in water (3 mL) is slowly addedto a solution of aniline 17b (1.5 g, 8.5 mmol) dissolved in AcOH (12 mL)and 2 M HCl (25 mL) at 0° C. The resulting mixture is stirred at thistemperature for 1 h. Solid CuCl (2.6 g, 26 mmol) is added portionwise(over 2 min) and the reaction is allowed to come to RT, is then stirredfor 30 min and then is warmed to 60° C. for 40 min. The mixture ispoured into water (100 mL) and extracted with EtOAc (2×100 mL). Theorganic layer is dried with MgSO₄, filtered and evaporated to dryness.The product is purified by CombiFlash® Companion to afford aryl chloride17c (1.11 g, 99% yield) as a pale yellow solid.

Step 3:

Solid pre-activated Zn dust is added to a solution of ketone 17c inAcOH. The reaction mixture is then heated to 100° C. and stirred at thattemperature for 4 h. The reaction mixture is filtered (EtOAc washing),the filtrate is evaporated to dryness and the product purified byCombiFlash® Companion to afford indane 17d (902 mg, 88% yield) as awhite crystalline solid.

Step 4:

A solution of BBr₃ in DCM (1 M, 9.9 mL, 9.9 mmol) is added dropwise to aprecooled (−78° C.) solution of methyl ether 17d (902 mg, 4.9 mmol)dissolved in DCM (20 mL). The reaction solution is stirred at thistemperature for 10 min and allowed to warm to RT. After stirring for 1.5h, water (50 mL) is added (caution! Exothermic!) and the mixture isextracted with DCM (3×50 mL). The combined organic layers are dried overMgSO₄, filtered and evaporated to dryness. The product is purified byCombiFlash® Companion to afford phenol 17e (700 mg, 84% yield) as anoff-white solid.

Step 5:

Tf₂O (1.05 mL, 12 mmol) is added to a precooled (0° C.) solution ofphenol 17e (700 mg, 4.1 mmol) and Et₃N (1.7 mL, 12 mmol) in DCM (20 mL).The resulting dark solution is allowed to warm to RT. After 25 min, thereaction is quenched with saturated NaHCO₃ (10 mL), diluted with DCM,and the organic layer washed with water, brine, dried over MgSO₄ andevaporated to dryness. The product is purified by CombiFlash® Companionto afford triflate 17f (1.21 g, 97% yield) as a yellow oil.

Step 6:

A solution of triflate 17f (1.2 g, 4.0 mmol), bis[pinocolato]diborane(1.5 g, 6.0 mmol) and potassium acetate (1.3 g, 14 mmol) in DMF (20 mL)is degassed with Ar for 5 min followed by the addition of thePdCl₂dppf-DCM complex (490 mg, 0.60 mmol). The reaction mixture is thendegassed for an additional 5 min before being heated to 95° C. for 5 h.The reaction is then cooled to RT. The crude reaction mixture is dilutedwith water and the product is extracted with EtOAc (3×100 mL). Thecombined organics are washed with water (100 mL) and brine (100 mL). Theorganic phase is then dried over MgSO₄ and filtered and concentrated.The crude mixture is further purified by CombiFlash® Companion to affordboronic ester 17g (593 mg, 53% yield) as a pale yellow solid.

Example 18 Synthesis of Boronate Fragment 18d (Used in Preparation of1067)

Step 1:

Neat Tf₂O (0.83 mL, 4.9 mmol) is added dropwise to a cooled (0° C.)solution of phenol 18a (0.50 g, 3.1 mmol) and pyridine (1.3 mL, 17 mmol)in DCM (15 mL). The reaction is allowed to warm to RT and stirovernight. The reaction is quenched by the addition of a 10% citric acidsolution (50 mL) and the mixture is extracted with DCM (3×50 mL). Thecombined organics are washed with water (50 mL), dried over MgSO₄,filtered and concentrated. The product is purified by CombiFlash®Companion to give triflate 18b (500 mg, 94% yield).

Step 2:

Deoxyfluor® (0.83 mL, 4.2 mmol) followed by EtOH (10 uL, 0.2 mmol) areadded to neat triflate 18b (500 mg, 1.7 mmol) in a sealable tube. Thetube is sealed and the reaction is heated in an oil bath at 85° C. andis stirred overnight. The reaction is then cooled to 0° C. and quenchedby the slow addition of NaHCO₃ (100 μL, CAUTION! Exothermic!). Themixture is diluted with water (50 mL) and extracted with DCM (3×50 mL).The combined organic layers are washed with water (50 mL) and brine (50mL). The organic phase is then dried over MgSO₄, filtered andconcentrated. The crude product is purified by CombiFlash® Companion toprovide the difluorotetrahydronaphtyl triflate 18c (175 mg, 33% yield).

Step 3:

Step three is performed exactly as in step 6 of Example 17 to provideboronic ester 18d.

Example 19 Synthesis of Boronate Fragment 19d (Used in Preparation of1070, 1078)

Step 1:

Solid N-chlorosuccinimide (2.2 g, 16 mmol) is added in portions over 5min to a solution of naphthylamine 19a (2.3 g, 16 mmol) dissolved inCCl₄ (150 mL). The reaction is then heated to 50° C. and is stirred for40 min. The reaction is then cooled to RT, solids are removed byfiltration and the filtrate is washed with water (100 mL), dried overMgSO₄ and evaporated to dryness to provide chloroaniline 19b (2.8 g, 96%yield).

Step 2:

A solution of NaNO₂ (1.2 g, 17 mmol) in water (5 mL) is slowly added toa precooled (0° C.) suspension of aniline 19b (2.8 g, 15 mmol) in 12 NHCl (7 mL) and ice (9.7 g), so as to maintain the temperature below 5°C. The mixture is stirred for 15 min and then is transferred to asolution of KI (8.7 g, 52 mmol) in water (30 mL) and the resultingmixture is stirred for 2 h. The mixture is extracted with Et₂O (3×100mL) and the combined organic layers washed successively with 3 N NaOH(2×50 mL), 5% NaHSO₃ (50 mL) and brine (100 mL). The organic phase isdried over MgSO₄, filtered and concentrated to dryness. The crudeproduct is purified by flash chromatography (EtOAc/hexanes) to providearyl iodide 19c (2.4 g, 54% yield).

Step 3:

Step three is carried out exactly as described in step 11 of Example 15to provide boronic ester 19d.

Example 20 Synthesis of Boronate Fragment 20d (Used in Preparation of1064)

Step 1:

Allyl bromide (2.1 mL, 25 mmol) followed by potassium carbonate (7.2 g,52 mmol) are added to a solution of 6-chlororesorcinol 20a (10 g, 69mmol) dissolved in DMF (120 mL). The reaction is stirred overnight,diluted with EtOAc (500 mL) and washed with water (3×500 mL). Theorganic layer is dried over MgSO₄ and concentrated to dryness. The crudeproduct is purified by CombiFlash® Companion to obtain allyl ether 20b(1.8 g, 40% yield).

Step 2:

Methyl iodide (1.2 mL, 20 mmol) followed by potassium carbonate (3.8 g,27 mmol) are added to a solution of phenol 20b (1.8 g, 9.8 mmol)dissolved in DMF (12 mL). The reaction is stirred for 2 h, diluted withEtOAc (50 mL) and washed with water (3×50 mL). The organic layer isdried over MgSO₄ and concentrated to dryness. The crude product ispurified by CombiFlash® Companion to obtain methyl ether 20c (1.8 g, 40%yield).

Step 3:

Step 3 is comprised of a sequence of steps identical to steps 2 through6 of Example 12, followed by step 1 of Example 13 to provide boronicester 20d.

Example 21 Synthesis of Boronate Fragment 21g (Used in Preparation of1071)

Step 1:

Solid CuBr₂ (7.9 g; 35 mmol) is added to a solution of 21a (4.0 g, 23mmol) dissolved in EtOAc (32 mL) and CHCl₃ (32 mL). The mixture isheated to reflux and is stirred for 8 h. CuBr₂ (3.9 g, mmol) is thenadded and the mixture continues to stir at reflux for an additional 15h. The mixture is cooled to RT, the solids removed by filtration (EtOAcwashing). The filtrate is concentrated to afford the crude bromoketone21b (6.3 g), which is used directly in the next step.

Step 2:

Solid KF (2.5 g, 43 mmol) is added to a solution of crude bromoketone21b (6.3 g, 23 mmol) dissolved in DMF (21 mL). The reaction is stirredat RT for 3 h and then taken up in ether (300 mL), washed with brine(3×100 mL), dried over MgSO₄, filtered and concentrated to dryness. Thecrude product is purified by CombiFlash® Companion to afford ether 21c(2.1 g, 49% yield over two steps).

Step 3:

Solid NaBH₄ (270 mg, 7.1 mmol) is added to a precooled (0° C.) solutionof ketone 21c (1.0 g, 5.9 mmol) dissolved in MeOH (20 mL). The reactionis allowed to stir for 1 h and then quenched with aqueous HCl (1 N, 1mL). The volatiles are removed in vacuo and the product extracted withEtOAc (20 mL). The organic layer is washed with brine (20 mL), dried(Na₂SO₄), filtered and concentrated to afford the crude alcohol 21d (1.0g), which is used directly in the next step.

Step 4:

Solid AgNO₃ (1.0 g, 6.1 mmol) followed by I₂ (1.6 g, 6.2 mmol) are addedto a solution of alcohol 21d (1.0 g, 6.2 mmol) dissolved in MeOH (58mL). The mixture is stirred at RT for 1 h and then a solution of Na₂S₂O₄(0.5 M, 10 mL) is added and the mixture is stirred for 30 min. The MeOHis removed in vacuo and the residue taken up in EtOAc (50 mL), washedwith water (50 mL), brine (50 mL), dried (Na₂SO₄), filtered andconcentrated to afford aryl iodide 21e (1.6 g), which is used directlyin the next step.

Step 5:

Crude alcohol 21e (1.6 g; ˜5 mmol) is dissolved in a mixture of DCM (20mL) and TFA (2.2 mL). The reaction is stirred for 45 min and thenconcentrated to dryness. The residue is taken up in EtOAc (50 mL),washed with saturated NaHCO₃ (50 mL) and brine (50 mL). The organiclayer is dried over Na₂SO₄, filtered and concentrated to dryness. Thecrude product is purified by CombiFlash® Companion to provide benzofuran21f (978 mg, 65% yield over 3 steps).

Step 6:

Step 6 is carried out exactly as described for step 11 of Example 15 toprovide boronic ester 21g.

Example 22 Synthesis of Boronate Fragment 22d (Used in Preparation of1068)

Step 1:

Neat 3-bromo-2-methylpropene (1.7 mL, 16 mmol) is added to a suspensionof phenol 22a (3.0 g, 14 mmol) and potassium carbonate (5.6 g, 41 mmol)in DMF (35 mL). The reaction is stirred for 2 h and then quenched withwater (100 mL) and extracted with hexanes (2×100 mL). The organic phaseis washed with brine (2×100 mL) and concentrated to give ether 22b (3.3g, 87% yield).

Step 2:

Neat tributyltin hydride (2.3 mL, 8.8 mmol) is added to a solution ofaryliodide 22b (2.0 g, 7.3 mmol) and AIBN (120 mg, 0.73 mmol) in PhMe(40 mL) and the reaction is then stirred at reflux under N₂. After 1 h,the reaction is concentrated to dryness and the crude product purifiedby CombiFlash® Companion to provide dihydrobenzofuran 22c (785 mg, 73%yield).

Step 3:

Step 3 is comprised of a sequence of synthetic steps identical to steps10 and 11 of Example 15 to provide boronic ester 22d.

Example 23 Synthesis of Boronate Fragment 23c (Used in Preparation of1040, 1041, 1057)

Step 1:

Neat Tf₂O (056 mL, 3.3 mmol) is added dropwise to a cooled (0° C.)solution of phenol 23a (350 mg, 2.1 mmol; prepared according to Doi etal Bull. Chem. Soc. Jpn. 2004 77, 2257-2263) and pyridine (0.91 mL, 11mmol) in DCM (10 mL) under an Ar atmosphere. The reaction is allowed towarm to RT and then is stirred for abut 2 h. The reaction is quenched bythe addition of a 10% citric acid solution (20 mL) and extracted withDCM (3×20 mL). The combined organic layers are washed with water (20mL), dried over MgSO₄, filtered and concentrated to dryness. The crudeproduct is purified by CombiFlash® Companion to provide triflate 23b(512 mg, 82% yield).

Step 2:

A solution of the triflate 23b (510 mg, 1.7 mmol),bis[pinocolato]diborane (560 mg, 2.2 mmol) and potassium acetate (500mg, 5.1 mmol) in DMF (18 mL) is degassed with Ar for 5 min followed bythe addition of the PdCl₂dppf-DCM complex (140 mg, 0.17 mmol). Thereaction mixture is then degassed for an additional 5 min before beingheated to 100° C. by microwave irradiation for 10 min. The reaction isthen cooled to RT. The crude reaction mixture is diluted with EtOAc (60mL) and washed with brine (3×60 mL). The organic layer is dried overMgSO₄, filtered and concentrated. The crude mixture is further purifiedby CombiFlash® Companion to afford boronic ester 23c (200 mg, 42%yield).

Example 24 Synthesis of Boronate Fragment 24b (Used in Preparation of1061)

Step 1:

Compound 24b is prepared from 24a following a synthetic sequenceidentical to steps 1 to 6 of Example 12.

Example 25 Synthesis of Boronate Fragment 25b (Used in Preparation of1059)

Step 1:

Compound 25b is prepared from 25a following a synthetic sequenceidentical to steps 1 to 6 of Example 12.

Example 26 Synthesis of Boronate Fragment 26b (Used in Preparation of1105, 1106)

Step 1:

Compound 26b is prepared from 26a following a synthetic sequenceidentical to steps 1 to 6 of Example 12.

Example 27 Synthesis of Boronate Fragment 27b (Used in Preparation of1033)

Step 1:

Compound 27b is prepared from 27a following a synthetic sequenceidentical to steps 1 to 6 of Example 14.

Example 28 Synthesis of Boronate Fragment 28b (Used in Preparation of1060)

Step 1:

Compound 28b is prepared from 28a following a synthetic sequenceidentical to steps 1 to 8 of Example 6.

Example 29 Synthesis of Boronate Fragment 29b (Used in Preparation of1052)

Step 1:

Compound 29b is prepared from 29a following a synthetic sequenceidentical to steps 1 to 6 of Example 14.

Example 30 Synthesis of Boronate Fragment 30b (Used in Preparation of1080)

Step 1:

Compound 30b is prepared from 30a following a synthetic sequenceidentical to steps 2 and 3 of Example 18.

Example 31 Synthesis of Boronate Fragment 31b (Used in Preparation of1013)

Step 1:

Compound 31b is prepared from 31a following a synthetic sequenceidentical to steps 9 to 11 of Example 15.

Example 32 Synthesis of Boronate Fragment 32b (Used in Preparation of1005)

Step 1:

Compound 32b is prepared from 32a following a synthetic sequenceidentical to steps 5 to 6 of Example 17.

Example 33 Synthesis of Boronate Fragment 33b (Used in Preparation of1023, 1034)

Step 1:

Compound 33b is prepared from 33a following a synthetic sequenceidentical to steps 1 and 3 of Example 11.

Example 34 Synthesis of Boronate Fragment 34f (Used in Preparation of1094)

Step 1:

Benzyl bromide (25 mL, 210 mmol) followed by potassium carbonate (44 g,320 mmol) are added to a solution of 2-methylresorcinol 34a (38 g, 310mmol) dissolved in DMF (1 L). The reaction is stirred overnight, dilutedwith EtOAc (2 L) and washed with water (3×2 L). The organic layer isdried over Na₂SO₄ and concentrated to dryness. The crude product ispurified by CombiFlash® Companion to obtain benzyl ether 34b (18.6 g,39% yield).

Step 2:

Allyl bromide (3.0 mL, 35 mmol) followed by potassium carbonate (6.5 g,47 mmol) are added to a solution of phenol 34b (5 g, 23 mmol) dissolvedin DMF (100 mL). The reaction is stirred overnight, diluted with EtOAc(500 mL) and washed with water (3×500 mL). The organic layer is driedover Na₂SO₄ and concentrated to dryness. The crude product is purifiedby CombiFlash® Companion to obtain benzyl ether 34c (4.4 g, 75% yield).

Step 3:

Compound 34d is prepared from 34c following a synthetic sequenceidentical to steps 2 to 4 of Example 12.

Step 4:

Benzyl ether 34d and Pd—C (10% w/w, 100 mg, 0.094 mmol) are combined inEtOAc (5 mL) and the flask is evacuated and backfilled with a H₂atmosphere (balloon). After stirring for 3 h, the reaction is filteredthrough Celite® (EtOAc washing) and the filtrated concentrated to givephenol 34e (145 mg, 95% yield).

Step 5:

Compound 34f is prepared from 34e following a synthetic sequenceidentical to steps 5 to 6 of Example 17.

Example 35 Synthesis of Boronate Fragment 35e (Used in Preparation of1047)

Steps 1 through 4 are done in analogy to steps 3 through 6 from Example17.

Example 36 Synthesis of Boronate Fragment 36d (Used in Preparation of1075, 1076)

Step 1:

4-bromo-3-nitrotoluene 36a (5.0 g, 22.9 mmol) is dissolved in 50 mLethyl acetate and solid tin(II) chloride dihydrate (20.0 g, 86.9 mmol)is added. The mixture is heated under nitrogen atmosphere at 70° C. for2 h (note: temporary overheating to 100° C. is observed! Caution shouldbe exercised!). The mixture is cooled down and is poured into 200 mL ofice-water. 50 mL of 5% aqueous NaHCO₃ solution is added (rapidfoaming!), followed by 10 N aqueous NaOH to bring the pH ˜7-8. Largevolume of gelatinous yellowish precipitate is formed. This heterogeneousmixture is shaken with EtOAc (200 mL) and the mixture is centrifuged in50 mL portions, resulting in good separation of a yellowish solid. Theclear supernatant is decanted and is extracted with EtOAc. Combinedorganic phase is washed with brine, dried over sodium sulphate, filteredand concentrated under vacuum to give an orange oily residue. Thisresidue is re-dissolved in 100 mL of ether and the solution is washedwith 10% Na₂CO₃ (20 mL) followed by 2.5 M aqueous NaOH (20 mL). The darkbrown organic solution is then stirred with MgSO₄ and active charcoaland filtered to give a light yellow solution, which darkened rapidly onstanding in open flask. The solvent is removed under vacuum to give thedesired compound 36b as a brown-red oil which is used in the next stepwithout further purification (3.31 g, 78% yield).

Step 2:

A mixture of compound 36b (3.3 g, 17.7 mmol), glycerin (3.3 g, 35.5mmol), nitrobenzene (2.2 g, 17.7 mmol) and 75% aqueous sulfuric acid (10mL, 138 mmol) is stirred at 150° C. for 3 h (mixture turns black andviscous). The reaction mixture is cooled down, poured into ice-water(200 mL) and 10 N aqueous NaOH is added (30 mL, 300 mmol). The blackmixture is then shaken with EtOAc (100 mL) and is centrifuged in 50 mLportions. The upper EtOAc layers are combined and the bottom aqueouslayers containing the black tar are shaken with EtOAc andre-centrifuged. All EtOAc extracts are combined, washed with brine,dried over Na₂SO₄, filtered and concentrated under vacuum to give 4.8 gof a brown-red oil. This material is chromatographed on 80 g silica gelcolumn (CombiFlash® Companion apparatus, hexanes-EtOAc gradient). Thefractions containing the compound are concentrated under vacuum toafford compound 36c as a white solid (3.26 g, 83% yield).

Step 3:

To a cooled (−78° C.) solution of compound 36c (500 mg, 2.25 mmole) inanhydrous Et₂O (20 mL), is added over 5 min under an Ar atmosphere a 1.6M solution of n-BuLi in hexane (3.5 mL, 5.60 mmol). The mixture isstirred at −78° C. for 50 min, triisopropylborate (2.00 mL, 8.55 mmol)is then added dropwise and the mixture is stirred for 2 h at thattemperature. The mixture is slowly allowed to reach RT over a 2 h periodand it is poured into 1 M aqueous HCl (30 mL). The mixture istransferred into a separatory funnel, the organic layer is separated andthe aqueous layer is washed with Et₂O. The aqueous layer is thentransferred into a 500 mL Erlenmeyer flask and the pH of the solution isadjusted to approximately 6.3 (measured with a pH meter) by slowlyadding a saturated solution of NaHCO₃ in water (˜25 mL, careful:foaming!). The suspension is filtered off and the separated light-beigesolid is washed with water and dried under high vacuum. This crudeproduct (383 mg) is triturated with Et₂O/hexanes to give a first crop ofthe desired compound 36d as a free base (120 mg, 28% yield). The motherliquors are concentrated under vacuum and are purified by reversed-phaseHPLC using a CH₃CN/H₂O gradient containing 0.06% TFA (ODS-AQ, C-18column, 75×30 mm, 5-μm particle size). After lyophilization, a secondcrop of compound 36d is obtained as a TFA salt (102 mg, 15% yield),(total yield: 43%).

Example 37 Synthesis of Boronate Fragment 37d (Used in Preparation of1084)

Step 1:

1-bromo-4-chloro-2-nitrobenzene 37a is transformed to compound 37b usingthe procedure of example 36b, except for the fact that Et₂O is used forthe extractions instead of EtOAc.

Step 2:

Compound 37b (4.2 g, 20.3 mmol) is melted at 50° C. in a 100 mLround-bottomed flask containing a stirring bar and immersed in an oilbath. A solution of zinc chloride (700 mg, 5.03 mmol) and ferricchloride (540 mg, 3.25 mmol) in water (3.3 mL) is added in one portionfollowed by absolute EtOH (20 mL). The flask is stoppered with a rubbersepta and a needle is inserted to avoid any pressure build-up. Themixture is warmed to 80° C. and acrolein (1.68 mL, 24.4 mmol) is addedvia a syringe pump over a 2 h period. After the addition, the mixture isstirred at 80° C. for 1 h and an additional amount of solid ferricchloride is added (4.1 g, 25.3 mmol). The mixture is stirred at 80° C.for an extra 24 h and then concentrated under vacuum to give asemi-solid residue. Water (200 mL) is added followed by a 10 N aqueoussolution of NaOH (20 mL) and CH₂Cl₂ (200 mL). After shaking the mixturefor a few min, the solid is filtered over a pad of Celite® and thefiltrate is transferred into a separatory funnel. The organic layer isseparated and the aqueous layer is extracted with CH₂Cl₂. The combinedorganic extracts are washed with brine, dried (Na₂SO₄), filtered andconcentrated under vacuum to give 3.69 g of a brown solid. This solid istriturated in hot CH₃CN and filtered. The solid is discarded and thefiltrate is concentrated under vacuum to give 2.3 g of a brownsemi-solid. This material is purified on a CombiFlash® Companionapparatus on 40 g silica gel column eluted with EtOAc/hexanes gradient.After evaporation of the solvent under vacuum, the desired compound 37cis isolated as a yellow solid (390 mg, 8% yield).

Step 3:

Compound 37c is transformed to compound 37d using the procedure ofexample 36d.

Example 38 Synthesis of Boronate Fragment 38c (Used in Preparation of1085)

Step 1:

2-bromoaniline 38a is transformed to compound 38b using the procedure ofexample 37c except that methyl vinyl ketone is used instead of acrolein.

Step 2:

Compound 38b is transformed to compound 38c using the procedure ofexample 36d.

Example 39 Synthesis of Boronate Fragment 39k (Used in Preparation of1131 and in Example 46)

Reference: Feliu, L.; Ajana, W.; Alvarez, M.; Joule, J. A. Tetrahedron1997, 53, 4511.

Step 1:

Meldrum's acid 39b (47.04 g, 326 mmol) is taken in trimethylorthoformate (360 mL) and refluxed for 2 h. Then 2,5-dimethoxy aniline39a (50 g, 326 mmol) is added and the mixture is refluxed for an extra 5h. The reaction mixture is cooled down to RT and the solid which formsupon cooling is collected by filtration. It is further crystallized fromMeOH to afford compound 39c as a yellow solid (63 g, 63% yield).

Step 2:

Compound 39c (62.00 g, 202 mmol) is dissolved in diphenyl ether (310 mL)and refluxed at 240° C. for 30 min. The mixture is then cooled down toRT and n-hexane is added, which causes a brown precipitate to form. Thissolid is separated by filtration and is washed with n-pentane andn-hexane to remove non-polar impurities and the remaining dark brownsolid (compound 39d) is used as is in the next step (27 g, 65% yield).

Step 3:

A mixture of compound 39d (30.0 g, 146 mmol), DMAP (3.75 g, 30.7 mmol)and 2,6-lutidine (24.4 mL; 208 mmol) in DCM (1.4 L) is cooled to 0° C.and Tf₂O (29.6 mL, 175 mmol) is added slowly at 0° C. The resultingmixture is stirred at 0° C. for 2 h and at RT for 1 h. It is thendiluted with DCM, washed with H₂O and brine and dried (Na₂SO₄). Thesolvent is removed under reduced pressure and the residue is purified byflash chromatography on silica gel (20% EtOAc/petroleum ether). Thedesired compound 39e is isolated as a yellow solid (35 g, 71% yield).

Step 4:

A mixture of diisopropylethyl amine (46.5 mL, 267 mmol) in dry DMF (250mL) is degassed with argon for 30 min and is added to a mixture ofcompound 39e (30.0 g, 88.5 mmol), triphenylphosphine (7.70 g, 29.4mmol), tris(dibenzylideneacetone)di-palladium(0)-chloroform adduct (9.21g, 8.9 mmol). The resulting mixture is stirred for 5 min at 0° C. andTMS acetylene (13.4 g, 136 mmol) is added dropwise. The temperature israised to RT and the mixture is stirred for 4 h. Diethyl ether and wateris added, the aqueous layer is separated and washed with diethyl ether.The combined organic layers are washed with H₂O and brine. After dryingon Na₂SO₄, the solvent is removed under reduced pressure and the residueis purified by flash chromatography on silica gel (30% EtOAc/petroleumether). Compound 39f is isolated as a yellow solid (18 g, 70% yield).

Step 5:

A solution of ceric ammonium nitrate (42.3 g, 77.2 mmol) in H₂O (47 mL)is added under argon atmosphere to a solution of compound 39f (11.0 g,38.3 mmol) in acetonitrile (366 mL). The reaction mixture is degassedwith argon for 10 min and the mixture is stirred at RT for 20 min. Wateris then added and the solution is extracted with CH₂Cl₂. The organicextracts are combined, washed with H₂O, brine and dried (Na₂SO₄). Thesolvent is removed under reduced pressure and the residue is purified byflash chromatography on silica gel (40% EtOAc/petroleum ether). Thedesired compound 39g is isolated as a yellow solid (5.0 g, 52% yield).

Step 6:

Compound 39g (1.80 g, 7.1 mmol) is taken in distilled acetic acid (72mL) under argon atmosphere. Ammonium chloride (7.55 g, 141 mmol) isadded and the reaction is refluxed for 45 min. The reaction mixture iscooled to RT, H₂O is added and the solution is washed with EtOAc. Theaqueous layer is neutralized with a saturated aqueous solution of NaHCO₃and is extracted with EtOAc. The combined organic extracts are washedwith H₂O, brine and dried (Na₂SO₄). The solvent is removed under reducedpressure to afford compound 39h as a brown solid (250 mg, 19% yield).

Step 7:

Compound 39h (230 mg, 1.24 mmol) is dissolved in absolute EtOH (11 mL)and 10% palladium on carbon is added (10% w/w, 23 mg) under nitrogenatmosphere. The mixture is stirred for 15 h under one atmosphere ofhydrogen. The reaction is degassed with nitrogen, filtered throughCelite®, and the Celite® bed is washed with an EtOH—CHCl₃ mixture. Thesolvent is removed under reduced pressure to give compound 39i as abrown sticky solid (200 mg, 86% yield).

Step 8:

Compound 39i (600 mg, 3.21 mmol) is taken in dry CH₂Cl₂ (30 mL) undernitrogen atmosphere. The solution is cooled to 0° C. and triethylamine(0.89 mL, 6.42 mmol) is added dropwise followed by Tf₂O (0.65 mL, 3.87mmol). The temperature is raised to RT and the reaction mixture isstirred for 2 h. The mixture is diluted with CH₂Cl₂ and is washed withH₂O, brine and dried (Na₂SO₄). The solvent is removed under reducedpressure to afford a residue which is purified by flash chromatography(10% EtOAc/hexanes). Compound 39j is isolated as a brown solid (630 mg,61% yield).

Step 9:

In a dry (oven-dried for 30 min) 5-mL glass microwave vessel containinga magnetic stirring bar, are added compounds 39j (250 mg, 0.78 mmol),bis(pinacolato)diboron (250 mg, 0.94 mmol), anhydrous potassium acetate(150 mg, 1.51 mmol), Pd(PCy₃)₂ (62.0 mg, 0.091 mmol) and anhydrous,deoxygenated (argon bubbling for 30 min) 1,4-dioxane (4 mL). The vial iscapped tightly with a septum-cap and the vessel is flushed with argon.The mixture is stirred at 95° C. (oil bath temperature) under anatmosphere of argon for 16 h. The reaction mixture is then concentratedunder vacuum, the brown oily residue is dissolved in 7 mL of glacialAcOH and is filtered via 45 μm membrane filter. The dark brown solutionis divided into 5×1.5 mL portions and is injected on an automaticpreparative reversed-phase HPLC-MS apparatus (CH₃CN/H₂O gradientcontaining 0.06% TFA, ODS-AQ, C-18 column, 50×19 mm, 5-μm particlesize). The collected fractions are lyophylized to give the desiredcompound 39k as a yellow amorphous solid (115 mg, 45% yield for the TFAsalt).

Example 40 Synthesis of Triflate Fragment 40e (Used in Example 47)

Step 1:

A solution of fuming HNO₃ (6.0 mL, 142 mmol) and concentrated H₂SO₄ (0.2mL, 3.8 mmol) in chloroform (150 mL) is added to a solution of 2-fluoro4-methyl phenol 40a (20.0 g, 159 mmol) in chloroform (100 mL). Theresulting mixture is stirred for 2 h at RT and is transferred in aseparatory funnel. The solution is washed with H₂O, brine, the organiclayer is dried (Na₂SO₄) and the solvent is evaporated under reducedpressure. The reddish crude solid is crystallized from aqueous ethanolto give compound 40b as a yellowish solid (17 g, 62% yield).

Step 2:

Compound 40b is transformed to compound 40c using the procedure ofexample 36b.

Step 3:

A mixture of concentrated sulfuric acid (12.5 mL, 235 mmol), water (10mL), glycerol (10.0 mL, 137 mmol) and sodium 3-nitrobenzenesulfonate(9.57 g, 42.5 mmol) is heated gently until everything dissolved.Compound 40c (5.0 g, 35.5 mmol) is then added slowly to the warm (60°C.) solution and the mixture is heated at reflux for 2 h (bathtemperature: 140° C.). The reaction mixture is then cooled to roomtemperature and poured into ice-water. The solution is brought to pH 6-7with an aqueous ammonia solution. A brown precipitate of compound 40dformed, it is collected by filtration and dried under high vacuum (5.0g, 79% yield). This compound is used in the next step without furtherpurification.

Step 4:

To a solution of compound 40d (5.0 g, 28.0 mmol) and triflic anhydride(5.25 mL, 31.0 mmol) in CH₂Cl₂ (150 mL) is added dropwise at 0° C. Et₃N(4.7 mL, 33.6 mmol). The resulting mixture is warmed to RT and stirredfor 3 h. The reaction mixture is diluted with CH₂Cl₂ (50 mL), thesolution is washed with 1N HCl, water, brine and it is dried (Na₂SO₄).The solvent is removed under vacuum to give a dark solid, which ispurified by flash chromatography (10% EtOAc/hexanes) to afford thedesired compound 40e as an off-white solid (6.0 g, 68% yield).

Example 41 Synthesis of Compound 1006

Step 1:

Solid Pd(PPh₃)₄ (9 mg, 0.008 mmol) and CuI (3 mg, 0.015 mmol) aresuccessively added to a solution of 11c (200 mg, 0.75 mmol) and alkyne2f (190 mg, 1.1 mmol) dissolved in DMF (0.46 mL) and diethylamine (2.3mL). The reaction mixture is stirred overnight at RT and thenconcentrated, diluted with EtOAc (10 mL) and successively washed withbrine, 1 N aqueous HCl and water (10 mL each). The organic layer isdried over Na₂SO₄, concentrated under reduced pressure and the residuepurified by CombiFlash® Companion to give alkyne 41a (126 mg, 46% yield)

Step 2:

Tf₂O (96 μL, 0.57 mmol) is added via syringe over the period of 1 min toa stirred mixture of acetanilide (77 mg, 0.57 mmol) and 2-chloropyridine(67 μL 0.71 mmol) in DCM (1.0 mL) at −78° C. After 5 min, the reactionflask is placed in an ice-water bath and is warmed to 0° C. Alkyne 41a(102 mg, 0.29 mmol) in DCM (1 mL) is added via syringe. The resultingsolution is allowed to warm to RT. After stirring for 30 min, Et₃N (1mL) is added and the mixture is partitioned between DCM (50 mL) andbrine (50 mL). The organic layer is washed with brine (50 mL), is driedover anhydrous Na₂SO₄ and concentrated. The residue is then purified byCombiFlash® Companion giving quinoline 41b (81 mg, 60% yield).

Step 3:

LiBH₄ in THF (2 M, 255 μL, 0.51 mmol) is added to a solution of ester41b (81 mg, 0.17 mmol) dissolved in THF (900 μL) and the reaction isstirred overnight at RT. Excess reagent is quenched with HCl (threedrops, lots of effervescence) and the mixture neutralized with saturatedNaHCO₃ (10 mL) and extracted with EtOAc (3×10 mL). The combined organiclayers are dried over anhydrous Na₂SO₄ and concentrated to give alcohol41c (38 mg, 57% yield).

Step 4:

Dess-Martin periodinane (46 mg, 0.11 mmol) is added to a solution ofalcohol 41c (33 mg, 0.084 mmol) dissolved in DCM (1 mL). After 2 h, thereaction is applied to a pad of SiO₂ (1.5×1 cm) and the product iseluted with 1:1 hexanes/EtOAc (20 mL). The filtrate is evaporated togive the crude aldehyde. The aldehyde is then dissolved in 2:2:1THF/H₂O/t-butanol (2.5 mL) and one drop of 2,3-dimethyl-2-butene (0.8mL, 1 M in THF) is added. NaClO₂ (62 mg, 0.68 mmol) and NaH₂PO₄ (51 mg,0.42 mmol) are added as solids to the solution and the reaction isstirred at RT. After 30 min, the reaction is diluted with H₂O (5 mL) andextracted with EtOAc (3×10 mL). The organic layer is dried overanhydrous MgSO₄ and concentrated. The residue is purified by preparativeHPLC to give compound 1006 (12 mg, 27% yield).

It would be apparent to those skilled in the art that the abovesynthetic protocols can also be used in the synthesis of otherinhibitors where either 11c is replaced by another aromatic halide inStep 1 and/or the acetanilide is replaced with another aryl-NH—CO—R², orheteroaryl-NH—CO—R² (R²═CH₃) in Step 2.

Example 42 Synthesis of Compound 1017

Step 1:

Quinoline 11 (260 mg, 0.62 mmol), boronic ester 14g (350 mg, 1.3 mmol)and Pd[P(t-Bu)₃]₂ (50 mg, 0.098 mmol) are dissolved in DMF (4.3 mL) in amicrowave vial and a solution of Na₂CO₃ (1.25 mL, 2 M, 2.5 mmol) isadded. The solution is degassed (Ar balloon) and the mixture is thensubmitted to microwave heating at 120° C. for 10 min. The crude reactionmixture is diluted with water (15 mL) and the product is extracted withEtOAc (15 mL). The organic layer is washed with water (2×15 mL) anddried over anhydrous Na₂SO₄, filtered and concentrated. The crudeproduct is then purified by CombiFlash® Companion (gradient of EtOAc inhexanes) to give quinoline 42a as a mixture of atropisomers (175 mg, 65%yield).

Step 2:

An aqueous solution of LiOH (4 mL, 1N, 4 mmol) is added to a solution ofesters 42a in THF (25 mL), MeOH (6 mL) and water (12 mL) at RT and thereaction is heated to 50° C. After 4 h, the reaction is evaporated to awhite slurry under reduced pressure, diluted with 1 N NaOH (5 mL) andextracted with EtOAc (2×25 mL). The water layer is then acidified to pH˜3 with 10% HCl and extracted with DCM (2×100 mL) and EtOAc (100 mL).The combined organic layers are dried over anhydrous Na₂SO₄ andconcentrated. The desired product is isolated after purification of thiscrude by preparative HPLC to give the desired pure atropisomer(diastereomer) 1017 (7.5 mg, 4.4% yield).

Example 43 Synthesis of Compound 1125

Step 1:

In two separate batches, DMF (15 mL) and distilled water (3.0 mL) areadded to two microwave vials each charged with boronate 9b (560 mg, 2.06mmol), iodoquinoline 1i (600 mg, 1.45 mmol), potassium carbonate (602mg, 4.35 mmol) and Pd(PPh₃)₄ (252 mg, 0.218 mmol). The vials are thensealed and heated in a microwave reactor (7 min, 140° C.). The resultingmixtures are cooled, pooled and extracted with EtOAc (200 mL) and washedwith half-saturated aqueous NaHCO₃ (200 mL) and brine (200 mL). Theextract is dried over MgSO₄, filtered and evaporated to a red syrupwhich is chromatographed over silica gel (EtOAc/hexanes) to afford thepure atropisomers 43a (160 mg, 13% yield) and 43b (175 mg, 14% yield) aspale yellow amorphous solids, as well as a sample consisting of amixture of atropisomers which is set aside for later separation (275 mg,22% yield).

Step 2:

A solution of anilines 43a/43b (mixture of atropisomers; 50 mg; 0.116mmol) in anhydrous acetonitrile (0.4 mL) is added to a stirred mixtureof copper (II) bromide (32 mg; 0.145 mmol) and tert-butyl nitrite (22μL; 0.19 mmol) in anhydrous acetonitrile (0.6 mL) at RT under an argonatmosphere. After 1 h, the reaction is quenched with 1.0 N HCl andextracted with EtOAc (20 mL) and washed with water (20 mL) and brine (20mL). The extract is dried over MgSO₄, filtered and evaporated to afforda mixture of aryl bromides 43c/43d as a green solid which is used assuch (51 mg; 89% yield).

Step 3:

Sodium hydroxide (1.0 N, 1.00 mL; 1.00 mmol) is added to a stirredsolution of the ester mixture 43c and 43d (51 mg; 0.103 mmol) in MeOH(1.5 mL) and THF (3 mL) and the reaction heated to 50° C. After 16 h,the solution is acidified with 1.0 N HCl to a pH of ˜4 and extractedwith DCM (20 mL). The extract is dried over MgSO₄, filtered andevaporated to solid which is diluted with acetic acid and acetonitrile(to a volume of 2 mL) and purified by preparative HPLC (0.1% TFAwater/acetonitrile). The relevant fractions are pooled and lyophilizedto yield the TFA salts of inhibitors 43e (13 mg; 27% yield) and 1125 (18mg; 37% yield) as white powders.

It would be obvious to those skilled in the art that intermediate 43bcould also be used, employing the same methods, to prepare otherinhibitors such as the para-chloro analogue 1112. The bromo derivative43d could also be transformed via Suzuki coupling to para-alkylderivatives such as 1127.

Example 44 Synthesis of Compound 1103

Step 1:

To a solution of 3-bromo-2-methylaniline 44a (0.77 g, 4.15 mmol) in MeCN(20 mL) is added β-propiolactone (435 μL, 6.2 mmol). The reactionmixture is heated at reflux for 16 h. The reaction is found to beincomplete so an equivalent amount of the lactone is added again and thereaction mixture is heated at reflux for 24 h. The solvent is evaporatedbefore the residue is taken up into EtOAc and washed with 1 N HCl (aq)and brine before being dried (MgSO₄), filtered and concentrated.Purification by CombiFlash® Companion (hexanes/EtOAc) gives intermediate44b as a white solid (606 mg, 57% yield).

Step 2:

Compound 44b (1.1 g, 4.3 mmol) is combined with polyphosphoric acid (40g) and heated at 100° C. for 22 h. The cooled mixture is diluted withEtOAc and ice before being basified to pH ˜8 with 10 N NaOH. The phasesare separated and the aqueous phase again extracted with EtOAc (3×). Thecombined organic phases are dried (MgSO₄) filtered and concentrated.Purification by CombiFlash® Companion (hexanes/EtOAc) gives the desiredketone 44c as a yellow solid (535 mg, 52% yield).

Step 3:

To a solution of ketone 44c (489 mg, 2.04 mmol) in DCE (20 mL) is addedzinc iodide (975 mg, 3.06 mmol) and sodium cyanoborohydride (960 mg,15.3 mmol). The mixture is heated at 85° C. for 1.5 h. The mixture iscooled to RT and diluted with EtOAc and a saturated solution of NH₄Cl(containing 10% by volume of 6 N HCl). The mixture is stirred for 30 minbefore the phases are separated. The organic phase is washed withsaturated brine and then dried (MgSO₄) filtered and concentrated. Pureintermediate 44d (232 mg, 50% yield) is isolated after purification ofthe crude by CombiFlash® Companion (hexanes/EtOAc).

Step 4:

To compound 44d (260 mg, 1.15 mmol) in anhydrous DMF (10 mL) is addedbis(pinacolato)borane (380 mg, 1.5 mmol) followed by potassium acetate(339 mg, 3.45 mmol). The mixture is degassed with Ar for 10 min beforethe catalyst PdCl₂(dppf)-CH₂Cl₂ complex (141 mg, 0.17 mmol) is added.The mixture is heated at 95° C. for 20 h before being cooled and dilutedwith EtOAc and water. The organic phase is washed with saturated brine(3×) before being dried (MgSO₄), filtered and concentrated. The crudematerial is purified by CombiFlash® Companion (hexanes/EtOAc) to givethe boronate 44e as a yellow solid (252 mg, 80% yield).

Step 5:

In a vessel suitable for microwave heating, quinoline 1i (62 mg, 0.15mmol), boronate 44e (50 mg, 0.18 mmol), potassium carbonate (62 mg, 0.45mmol) and Pd[(PPh₃)]₄ (26 mg, 0.023 mmol) in DMF (2.5 mL) and water(0.25 mL) are added. The mixture is irradiated at 110° C. for 15 min ina microwave before being cooled and diluted with EtOAc. The organicphase is washed with brine (3×) before being dried (MgSO₄), filtered,concentrated and purified by combiflash (hexanes/EtOAc) to obtain amixture of atropisomers (diastereomers) as a yellow oil. This material(68 mg, 0.16 mmol) is dissolved in THF (1.5 mL) and MeOH (0.5 mL) beforebeing treated with 5 N NaOH (0.32 mL, 1.57 mmol, 10 eq). The mixture isheated at 50° C. for 18 h before being cooled. The pH is adjusted to −5with aqueous 1 N HCl and the mixture is extracted with EtOAc. Theorganic phase is washed with saturated brine before being dried (MgSO₄),filtered and concentrated. The atropisomers (diastereomers) areseparated by preparative HPLC to give the desired compound 1103 as alight orange solid (16.8 mg, 20% yield over two steps).

Example 45 Synthesis of a Compound 1074 Via Decarboxylative Biaryl CrossCoupling Reactions

The decarboxylative cross coupling reactions are used to prepare avariety of inhibitors; the details of the synthetic methodology can befound in the literature reference: J. Am. Chem. Soc. 2006, 128,11350-11351. An example is shown below:

In a vial suitable for microwave reactions is added5,6-dihydro-4H-cyclopenta[b]thiophene-2-carboxylic acid (73 mg, 0.44mmol), 4-iodoquinoline 1i (100 mg, 0.24 mmol), tetrabutylammoniumchloride hydrate (67 mg, 0.24 mmol), cesium carbonate (118 mg, 0.36mmol), and the catalyst Pd[(PtBu)₃]₂ (12.4 mg, 0.02 mmol) in DMF (3 mL).The vial is then capped and submitted directly to the microwaveconditions: 170° C. for 8 min. After cooling, the reaction is dilutedwith EtOAc (100 mL) and the mixture is washed with brine (3×), water(1×), before being dried (MgSO₄), filtered and concentrated. The residueis purified by CombiFlash® Companion (hexanes/EtOAc) to afford themethyl ester of the desired product (79 mg, 80% yield) as a foamy solid.The final compound 1074 is obtained after a saponification step followedby HPLC purification.

Example 46 Synthesis of Compound 1131

In a 5-mL glass microwave vessel containing a magnetic stirring bar, areadded compound 46a (100 mg, 0.234 mmol), compound 39k (90 mg, 0.273mmol), anhydrous potassium carbonate (150 mg, 1.08 mmol), Pd(PPh₃)₄(40.0 mg, 0.035 mmol), anhydrous, deoxygenated (Argon bubbling for 30min) dimethylacetamide (3 mL) and deoxygenated H₂O (0.35 mL). The vialis capped and is heated in a microwave at 100° C. for 25 min (BiotageInitiator apparatus). The mixture is cooled and THF (3 mL), H₂O (1 mL),and MeOH (3 mL) are added followed by a 10 N aqueous solution of NaOH(0.50 mL, 5.0 mmol). The reaction mixture is heated to 60° C. 1 h. Thereaction is cooled and the volatiles are removed under vacuum to give abrown oily residue which is diluted with 7 mL of acetic acid, filteredon a 45 μm membrane filter and injected in 1.5-mL batches into apreparative reversed-phase HPLC-MS for purification (CH₃CN/H₂O gradientcontaining 0.06% TFA, ODS-AQ, C-18 column, 50×19 mm, 5-μm particlesize). The desired atropisomer is re-purified under the same conditionsas described above. 1131 as a white amorphous solid (52.0 mg, 32% yield,bis-TFA salt).

Example 47 Synthesis of Compound 1101

In a dry pressure tube, compound 1i (100 mg, 0.24 mmol) is dissolved inanhydrous, deoxygenated THF (2.5 mL, Ar bubbling for 30 min) and themixture is cooled to −40° C. under Ar atmosphere. A freshly titratedsolution of i-PrMgCl—LiCl complex in THF (the titration is donefollowing the protocol in Lin, H. S.; Paquette, L. A. Synth. Commun.1994, 24, 2503; 0.83 M solution, 0.400 mL, 0.328 mmol) is added and isstirred at −40° C. for 30 min.

In a separate flask, a 0.65M solution of zinc chloride in THF isprepared in the following manner: 115 mg (0.84 mmole) of anhydrous zincchloride is placed in an oven-dried 2 mL glass microwave vessel and isdried at 180° C. (oil bath) under high vacuum overnight. The vessel iscooled to room temperature and 1.3 mL of anhydrous, argon-degassed THFis added. The mixture is sonicated until all the zinc chloridedissolved.

The 0.65M solution of zinc chloride in THF (0.50 mL, 0.30 mmol), isadded to the reaction mixture at −40° C. It is stirred at thistemperature for 5 min and is warmed to −5° C. and kept at thistemperature for 1 h and finally at RT for an extra hour. Pd₂(dba)₃ (24.0mg, 0 0262 mmol) and RuPhos (24.0 mg, 0.051 mmol, Strem Chemicals) areadded under an Ar atmosphere and the mixture is stirred at RT for 5 min.Compound 40e (80.0 mg, 0.259 mmol) is then added, the reaction vessel ispurged with Ar, sealed and heated on an oil bath set at 80° C. for 40 h.

The reaction mixture is cooled down and H₂O (0.30 mL), MeOH (0.30 mL)and 10 N aqueous NaOH (0.30 mL, 3.0 mmol) are added sequentially and themixture is heated to 60° C. for 2 h. Acetic acid is then added (2 mL),the mixture is filtered over a 45 μm membrane filter and the compound ispurified in two portions through direct injection into asemi-preparative reversed-phase HPLC-MS (CH₃CN/H₂O gradient containing0.06% TFA, ODS-AQ, C-18 column, 75×30 mm, 5-μm particle size). A mixtureof two atropisomers is isolated (20 mg, 13% yield for bis-TFA salt).Both atropisomers are separated using silica-gel chromatography(CombiFlash® Companion apparatus, 4 g column, MeOH—CH₂Cl₂ gradient) togive compound 1101 (5.5 mg, 5% yield) as a pure atropisomer.

Example 48 Synthesis of Boronate Fragment 48b (Used for the Preparationof 1136)

Step 1:

A stirred DMF (5 mL) solution of the arylbromide 48a (0.152 g, 0.71mmol), potassium acetate (0.209 g, 2.1 mmol) and bis(pinacolato)diborane(0.234 g, 0.92 mmol) is degassed by bubbling Ar through the solution for20 min. PdCl₂(dppf)-DCM (87 mg, 0.11 mmol) is added and degassing iscontinued for 15 min. The system is sealed (Teflon screw cap vessel)under Ar and heated to 90° C. for 16 h. The reaction mixture is allowedto cool to RT, dilute with EtOAc (150 mL), washed with brine (3×100 mL)and water (2×100 mL), dried over anhydrous MgSO₄, filtered andconcentrated to dryness. The residue is purified by CombiFlash®Companion (EtOAc/hexanes) to give the desired boronate 48b (144 mg, 77%yield) as a yellowish solid.

Example 49 Alternative Synthesis of Boronate Fragment 39k (Used for thePreparation of 1143, 1144, 1150, 1151, 1152, 1153)

Step 1:

1,3-acetonedicarboxylic acid 49a (30 g, 205.3 mmol) is added in portionsto acetic anhydride (55 g, 587.7 mmol) and the mixture is stirred at 35°C. for 23 h. The mixture is filtered and the filtrate is diluted withbenzene (200 mL) and the solution stored at 5° C. for 3 h. Theprecipitate that formed is filtered and dried under vacuum to givecompound 49b as a pale yellow solid (26.9 g, 70% yield).

Step 2:

To a stirred solution of aniline 49c (7.5 g, 44 mmol) in AcOH (50 mL) isadded 49b (8.0 g, 40 mmol) portionwise. Following addition, the reactionmixture is warmed to 35° C. After 2 h, the reaction mixture is cooled toroom temperature and poured in ice/water (600 mL). The resultingprecipitate is isolated by filtration, rinsed with water (100 mL) anddried under vacuum to give 49d (9.1 g, 61% yield).

Step 3:

Compound 49d (5.7 g, 15.4 mmol) is added portionwise to concentratedsulfuric acid (20 mL) at RT, temperature of the reaction mixture is keptbelow 30° C. during addition. The mixture is stirred at RT for 30 minand then poured in ice/water (400 mL). The resulting precipitate isisolated by filtration, rinsed with water and dried under vacuum to give49e (3.5 g, 72% yield) as a white solid.

Step 4:

The borane solution (1.0 M in THF, 10.5 ml, 10.5 mmol) is added dropwiseto an ice cold solution of quinolone 49e (1.5 g, 4.8 mmol) in dry THF(40 mL) under a N₂ atmosphere. After the addition, the reaction isallowed to warm to RT and stirred for 22 h (reaction not completed byHPLC, 15% starting material). An extra equivalent of BH₃ is added at 0°C. and the reaction mixture is heated to 45° C. for 2 h. The reactionmixture is carefully quenched with 1.0 N NaOH (10 mL) and THF is removedunder vacuum. The mixture is poured in EtOAc (100 mL) and the desiredcompound crashed out of the solution under these conditions. The solid49f is filtered and dried under vacuum (1.1 g, 79% yield) as grey solid.

Step 5:

To a solution of 49f (1.1 g, 3.8 mmol) in DCM (60 mL) at −78° C. isadded dropwise a 1.0 M BBr₃ solution (23 mL, 23 mmol). The cooling bathis removed after 1 h and the mixture is stirred at RT for 16 h (by HPLC,˜30% cyclized product 49h is formed). The mixture is poured in ice/water(100 mL) and the white precipitate that formed is filtered and driedunder vacuum to give 49g (773 mg, 71% yield).

Step 6:

To a solution of compound 49g (773 mg, 2.27 mmol) in THF (30 mL) isadded PPh₃ (928 mg, 3.5 mmol) followed by DIAD (0.69 ml, 3.5 mmol)(dropwise) and the solution is stirred at RT for 2 h. The reactionmixture is concentrated under vacuum and the crude product is directlyadded portionwise to POCl₃ (2 mL) at RT. The reaction mixture is stirredat 100° C. for 45 min and then cooled to RT. The mixture is concentratedunder vacuum (to remove POCl₃) and the crude product is diluted withDCM. The organic phase is washed with 1.0 N NaOH, water, and brine,dried (MgSO₄), filtered and concentrated under vacuum. The crude productis purified in two batches by combi-flash (330 column hexanes/EtOAc 9/1to 1/1) to give 49h as a pale yellow solid (445 mg, 91% yield).

Step 7:

To a solution of chloroquinoline 49h (30 mg, 0.1 mmol) in TFA (1 mL) isadded zinc (34 mg, 0.5 mmol). The reaction mixture is stirred at RT for16 h. The mixture is filtered, concentrated under vacuum, then dilutedwith 1.0 N NaOH (5 mL) and extracted with DCM (3×). The combined organicextracts are washed with water and brine, dried (MgSO₄), filtered andconcentrated under vacuum. The crude product was purified by combi flash(hexanes/EtOAc 6/4 to 4/6) to give 49i as a pale yellow solid (26 mg,quantitative yield).

Step 8:

The reaction is done following a procedure similar to the one in step 9of example 39 using Pd(PPh₃)₄ as catalyst and starting with 49i to give39k as a white solid.

Example 50 Synthesis of Boronate Fragment 50d (Used for the Preparationof 1018, 1020)

Step 1:

Solid NaBH₄ (603 mg, 15.9 mmol) is added to a solution of ketone 50a(4.11 g, 19.92 mmol) dissolved in MeOH (62 mL) at 0° C. The reaction iswarmed to RT and is allowed to stir for 2 h. The reaction is quenchedwith aqueous HCl (1 N, 20 mL), the MeOH is removed by concentration andthe product extracted with EtOAc (2×50 mL). The organic layer is washedwith brine (50 mL), dried over MgSO₄, filtered and concentrated toafford alcohol 50b (4.1 g, 97% yield). This material is used as is inthe next step.

Step 2:

To a cold solution (0° C.) of 50b (3.96 g, 19.31 mmol) in DCM (12 mL) isadded diethylamino sulfur trifluoride (2.78 mL, 21.25 mmol). Thereaction is warmed to RT and is allowed to stir for 2 h. The reaction isquenched with aqueous NaHCO₃ and extracted with DCM. The organic layeris dried with MgSO₄, filtered and evaporated to dryness. The product ispurified by CombiFlash® Companion to afford 50c (2.1 g, 52% yield) as acolorless oil.

Step 3:

Step 3 is carried out exactly as in step 1 of example 48 to provideboronic ester 50d.

Example 51 Synthesis of Boronate Fragment 51a (Used for the Preparationof 1115)

Step 1:

To a cooled solution (0° C.) of boronate 5f (400 mg, 1.45 mmol) inanhydrous DMF (8 mL) is added NaH (87.4 mg, 2.18 mmol, 60% dispersion inoil). The mixture is stirred for 30 min before being treated withiodoethane (233 μL, 2.9 mmol). The resultant mixture is stirred for 18 hbefore being quenched with water and extracted with EtOAc. The organicphase is washed with brine and dried over MgSO₄, filtered andconcentrated. The residue is purified by CombiFlash® Companion(EtOAc/hexanes) to give 51a as colourless oil (317 mg, 72%).

Example 52 Synthesis of Boronate Fragment 52g (Used for the Preparationof 1149)

Step 1:

To a solution of 4-bromo-1-methoxy-2-nitro-benzene 52a (6 g, 25.9 mmol)in dry THF (250 mL) at −40° C. is added dropwise the vinylmagnesiumbromide solution (1M in THF, 90.5 mL, 90.5 mmol). The reaction mixtureis stirred at −40° C. for 4 h then poured into saturated NH₄Cl solution.The reaction mixture is extracted with Et₂O (2×); the combined organiclayers are washed with brine, dried over Na₂SO₄, filtered andconcentrated in vacuo. The crude material was purified by flashchromatography eluting EtOAc/hexanes (10% to 40%) affording 52b (620 mg,11% yield).

Step 2:

A solution of indole 52b (619 mg, 2.7 mmol) in DMF (5 mL) at 23° C. istreated with NaH (60% in oil dispersion, 125 mg, 5.2 mmol) and stirredat 23° C. for 5 min. Ethyl bromoacetate (637 μL, 5.75 mmol) is added andthe solution is stirred at 23° C. for 24 h; HPLC/MS analysis indicated62% conversion. To this mixture, additional amounts of NaH (60% in oildispersion, 77 mg, 1.9 mmol) and ethyl bromoacetate (244 μL, 2.2 mmol)are added. After 10 min, the HPLC analysis indicated >90% conversion.The reaction is diluted with EtOAc, washed with saturated NH₄Cl solutionand brine (4×), dried over MgSO₄, filtrated, concentrated and purifiedby flash chromatography (5-20%; EtOAc/hexanes) to give 52c (541 mg, 63%yield) as a yellow oil.

Step 3:

To a solution of 52c (385 mg, 1.2 mmol) in THF (12.3 mL) is added LiBH₄solution (2 M in THF, 1.54 mL, 3.1 mmol). The mixture is stirred at RTfor 16 h, then cooled to 0° C. and neutralized with NH₄Cl (sat). Theresulting solution is extracted with ethyl acetate (2×) and combinedorganic layers are washed with water, brine, dried over Na₂SO₄, filteredand concentrated in vacuo providing the compound 52d (317 mg, 95%yield).

Step 4:

To a solution of 52d (284 mg, 1.05 mmol) in DCM (9.7 mL) at RT is addedthe AlCl₃ (561 mg, 4.2 mmol). This mixture is stirred at RT for 16 h,then cooled to 0° C. and methanol is added. The resulting solution isconcentrated in vacuo. The residue is diluted with DCM containing 5%MeOH and a mixture of brine/water 50:50. The resulting solution isextracted with DCM until no more products remained in the aqueous layer.The organic layer is dried over Na₂SO₄, filtered and concentrated invacuo. Purification by flash chromatography (2-10%, methanol/DCM) givesthe compound 52e (200 mg, 74% yield).

Step 5:

To a solution of 52e (170 mg, 0.66 mmol) in DCM (10 mL) at 0° C. isadded the triethylamine (204 μL, 1.46 mmol). Methanesulfonyl chloride(62 μL, 0.8 mmol) is added and the mixture is stirred at 0° C. for 10min. The reaction is not completed, additional methanesulfonyl chloride(30 μL, 0.4 mmol) is added and the mixture is stirred at 0° C. for 10min. Poured into ice water the extracted with CH₃Cl (3×). Combinedorganic layers are washed with saturated NH₄Cl, NaHCO₃, brine, driedover Na₂SO₄, filtered and concentrated in vacuo. The crude intermediate(261 mg, 0.78 mmol) is diluted in DMF (5.3 mL) and NaH (60% in oildispersion, 52.9 mg, 1.3 mmol) is added at 0° C. The mixture is stirredat RT for 16 h. Et₂O and brine are added, layers separated and theorganic layer is washed with brine (2×), dried over Na₂SO₄, filtered andconcentrated in vacuo. The product is then purified by flashchromatography eluting EtOAc/hexanes (10-25%) to give 52f (170 mg, 91%yield).

Step 6:

Step 6 is carried out exactly as in step 1 of example 48 to provideboronic ester 52g.

Example 53 Synthesis of Boronate Fragment 53i (Used for the Preparationof 1141, 1148)

Step 1:

To a solution of 53a (10.0 g, 48.3 mmol) in acetic acid (150 mL) at RTis added iron (10.8 g, 193 mmol) and the reaction mixture is stirred at70° C. for 2 h. The cooled reaction mixture is filtered and the filtrateconcentrated under vacuum. The residue diluted with EtOAc (300 mL) iswashed with water (100 mL), brine (100 mL), dried over MgSO₄, filteredand concentrated under vacuum to give 53b as a yellow solid (8.8 g, 100%yield). This material is used in the next step without furtherpurification.

Step 2:

To a solution of aniline 53b (8.8 g, 49.7 mmol) anddiethylphosphonoacetic acid (8.8 mL, 54.6 mmol) in DCM (300 mL) is addedHATU (22.7 g, 59.6 mmol) followed by DIPEA (21.6 mL, 124 mmol). Thereaction mixture is stirred at RT for 3 h. After that period, thetransformation is incomplete, therefore, more diethylphosphonoaceticacid (4.0 mL, 24.9 mmol), HATU (9.4 g, 24.9 mmol) and DIPEA (4.3 mL,24.9 mmol) are added. The mixture is stirred for another 2 h. Themixture is diluted with DCM (300 mL), washed with aqueous 0.2N HCl(3×100 mL), aqueous 0.2N NaOH (3×100 mL), water (100 mL) and brine (100mL). The organic phase is dried over MgSO₄, filtered and concentratedunder vacuum. The residue is dried under vacuum overnight to give thedesired product 53c as a pale orange solid (16.0 g, 60% yield). Thismaterial is used in the next step without further purification.

Step 3:

To a solution of 53c (16.0 g, 26.9 mmol) in THF (180 mL) at 50° C. iscarefully added NaH (60% in oil, 1.2 g, 29.6 mmol) and the reactionmixture is stirred for 2 h. The cooled reaction mixture is quenched withMeOH (20 mL) and silica gel (50 g) is added. The mixture is concentratedunder vacuum and purified by CombiFlash® Companion (DCM/MeOH) to givethe desired intermediate 53d (1.8 g, 27% yield).

Step 4:

A solution of 53d (1.8 g, 7.1 mmol) in POCl₃ (30 mL, 322 mmol) isstirred at 110° C. for 45 min. The cooled reaction mixture isconcentrated under vacuum. The residue is diluted with DCM (100 mL),washed with aqueous 1.0N NaOH (50 mL), water (50 mL), and brine (50 mL),dried over MgSO₄, filtered and concentrated under vacuum. The crudeproduct is purified by CombiFlash® Companion (DCM/MeOH) to give thedesired chloroquinoline 53e (1.3 g, 81% yield).

Step 5:

A solution of 53e (1.3 g, 5.8 mmol) in EtOH (100 mL) is degassed bybubbling argon for 45 min. Palladium (10 wt. % on activated carbon, 1.0g) is added to the solution and the reaction mixture is stirred under anatmospheric pressure of hydrogen for 6 h. The reaction mixture isfiltered and the filtrate concentrated under vacuum to give 53f (1.1 g,100% yield). This material is used in the next step without furtherpurification.

Step 6:

To a solution of 53f (1.1 g, 5.8 mmol) in DCM (40 mL) at 0° C. is addeda BBr₃ solution (1M in heptane, 12.7 mL, 12.7 mmol). The reactionmixture is stirred for 30 min at 0° C. and then slowly allowed to warmto Rt and stirred at this temperature for 24 h. The reaction mixture isquenched with MeOH (10 mL) and neutralized with aqueous 1.0 N NaOH. Theyellow precipitate that formed is filtered and dried under vacuum togive the desired product 53g (984 mg, 100% yield). This material is usedin the next step without further purification.

Step 7:

To a solution of 53g (984 mg, 5.7 mmol) and Et₃N (4.0 mL, 28.7 mmol) inDCM (40 mL) cooled to −78° C. is added Tf₂O) (2.1 mL, 12.6 mmol). Theresulting dark solution is stirred for 15 min at −78° C. and then slowlyallowed to warm to RT and then stirred at this temperature for 3 h. Thereaction mixture is diluted with DCM (50 mL), washed with aqueous 0.2 NHCl (25 mL), aqueous saturated NaHCO₃ (25 mL), water (25 mL), brine (25mL), dried over MgSO₄, filtered and concentrated under vacuum. Theresidue is purified by CombiFlash® Companion (DCM/MeOH) to give triflate53h (755 mg, 43% yield).

Step 8:

A well stirred DMF (7 mL) solution of triflate 53h (555 mg, 1.8 mmol),potassium acetate (608 mg, 6.4 mmol) and bis(pinacolato)diborane (697mg, 2.7 mmol) is degassed by bubbling argon through the solution for 20min. PdCl₂(dppf)-DCM (224 mg, 0.27 mmol) is added and degassing iscontinued for 15 min. The system is sealed (Teflon screw cap vessel)under argon and heated to 95° C. for 7 h. The mixture is poured inaqueous 1N HCl (30 mL) and diluted with EtOAc (15 mL). The layers areseparated and the aqueous layer is neutralized to pH 7 with aqueous 1.0N NaOH. This neutral aqueous layer is extracted with EtOAc (3×25 mL).The combined organic extracts are washed with brine (25 mL), dried overMgSO₄, filtered and concentrated under vacuum. The boronic acid 53i (290mg, 80% yield) is used as such for the following step.

Example 54 Synthesis of Boronate Fragment 54b (Used for the Preparationof 1145)

Step 1:

Compound 54b is prepared from 54a following a synthetic sequenceidentical to steps 1 to 8 of Example 53.

Example 55 Synthesis of Boronate Fragment 55g (Used for the Preparationof 1147)

Step 1:

To a 1L flask equipped with a mechanical stirrer is added 55a (30 g, 129mmol) in acetic acid (300 mL). To this mixture is added iron powder(14.4 g, 258 mmol) slowly in aliquots. The mixture is heated at 50° C.for 2 h before an additional 7.2 g (129 mmol) of iron powder is slowlyadded. After 1.5 h at 50° C., the conversion to the aniline is complete.The solution is cooled to RT and diluted with 500 mL of EtOAc beforebeing filtered through Celite. The filtrate is concentrated and thecrude product is partitioned between EtOAc (1 L) and 200 mL of water.The mixture is vigorously shaken and the organic phase is washed with200 mL of brine, dried over MgSO₄, filtered and concentrated to giveaniline 55b (23.7 g, 91%) as a brown oil which is used directly in thenext step.

Step 2:

Aniline 55b (600 mg, 3 mmol) is treated with 6N HCl (8 mL) and sonicateduntil a white suspension appears (HCl salt). This mixture is heated to100° C. before being treated with the vinyl ketone 55c (1.2 g, 5.9 mmol)which is prepared similarly to that reported (Ref: Bull. Korean Chem.Soc. 24 (2003) 1, 13-14) but via the Weinreb amide. The reaction mixtureis heated at reflux for 6 h, then stirred at RT for 16 h. The mixture isdiluted with EtOAc and basified with 10N NaOH. The organic phase isseparated and the aqueous phase re-extracted with EtOAc. The combinedorganic layers are washed with brine, dried (MgSO₄), filtered andconcentrated. The material is purified by Combiflash® Companion(EtOAc/hexanes) to afford the desired alcohol 55d (330 mg, 37.5%) as apale brown solid.

Step 3:

BBr₃ (3.34 mL, 3.34 mmol, 1M solution in DCM) is added to a solution ofalcohol 55d (330 mg, 1.12 mmol) in DCM (10 mL) at RT. The reactionmixture is stirred at RT for 16 h, then is quenched with MeOH andconcentrated to dryness. The residue is taken up into DCM and washedwith water and brine before being dried (MgSO₄) filtered andconcentrated to afford the bromophenol 55e (384 mg, 100%). This materialis used in the next step.

Step 4:

a mixture of bromophenol 55e (345 mg, 1.14 mmol) and K₂CO₃ (315 mg, 2.28mmol) in MeCN (20 mL) is treated at RT for 2 h. The mixture is thenconcentrated to dryness and the residue dissolved in EtOAc before beingwashed with water and brine. The organic phase is dried (MgSO₄),filtered and concentrated before the residue is purified by Combiflash®Companion (EtOAc/hexanes) to afford the cyclic ether 55f (301 mg, 60%)as an amber solid.

Step 5:

In a screw-cap pressure tube is added the cyclic ether 55f (180 mg, 0.68mmol), bispinocolatoborane (260 mg, 1.0 mmol), potassium acetate (226mg, 2.4 mmol) and Pd(dppf)Cl₂.DCM complex (83 mg, 0.10 mmol) in dry DMF(4 mL). The resulting mixture is de-gassed with Argon (5 min). The tubeis sealed and stirred at 95° C. for 2.5 h. The mixture is treated with1N HCl (10 mL) before being diluted with EtOAc. After separation of thelayers, the aqueous phase was neutralized with 1N NaOH (using a pHmeter) to pH 7.0 and is extracted with EtOAc (3×). The combined organiclayers are washed with brine, dried (MgSO₄), filtered and concentratedto afford the boronic acid 55g (124 mg, 79%). This material is used asis in the final cross coupling reaction.

Biological Data

The compounds of the invention have valuable pharmacological properties.Compounds from this class strongly associate with the integrase targetas demonstrated by an integrase displacement assay, are particularlyeffective at inhibiting HIV integrase and additionally show unexpectedpotency across at least four or all six major HIV-1 virus HIV variantsat residues 124, 125.

Integrase Displacement Assay A displacement assay is used to evaluatethe relative affinity of compounds of the invention to bind reversiblywith HIV integrase. The displacement assay measures the extent to whichProbe I (consisting of a HIV integrase inhibitor connected by a flexiblelinker to a biotin molecule), in complex with a His-tagged HIVintegrase, is displaced by compounds of the invention. Theintegrase-Probe I complex is formed in the presence or absence ofinhibitor compounds and the interaction is monitored by a homogeneoustime resolved fluorescence (hTRF) assay system.

Example 56 Production of Probe I

Step 1:

Dimethyl carbonate (22 mL, 269 mmol) and NaH (60% in oil, 10.8 g, 270mmol) are combined in toluene (80 mL) and heated to 90° C. for 20 min,then 4-chloroacetophenone P1 (14 mL, 109 mmol) is added dropwise overca. 15 min. The mixture is stirred at 90° C. for 30 min, then cooled andcarefully treated with 5% HCl (aq) (100 mL) and EtOAc (100 mL). Theorganic phase is washed with brine, dried (MgSO₄), filtered andconcentrated to dryness. The residue is purified by flash chromatography(15% EtOAc/hexanes) to give compound P2.

Step 2:

A mixture of compound P2 (7.1 g, 33.4 mmol) and 4-chloroaniline (5.9 g,46.3 mmol) in DMF/xylene (7 mL/40 mL) is heated at 140° C. for 10 h. Thecooled mixture is partitioned between 1M HCl (40 mL) and EtOAc (150 mL).The organic layer is washed with 1M HCl, water and brine, dried overMgSO₄, filtered and concentrated to dryness. The residue is purified byflash chromatography (SiO₂, 15% to 20% EtOAc/hexanes) to give compoundP3.

Step 3:

To a mixture of compound P3 (4.79 g; 15.5 mmol), KOtBu (2.0 g; 18.57mmol) and DMF (23 mL) is added ethyl-2-bromovalerate (3.2 mL, 18.25mmol). The mixture is allowed to stir at RT for 16 h, then is pouredover ice into a solution of 1N HCl (100 mL) and the mixture is extractedwith EtOAc (2×100 mL). The combined organic extracts are washed withbrine (4×), dried (Na₂SO₄), filtered and concentrated to afford, afterpurification by chromatography (EtOAc/hexanes) compound P4 as a mixtureof diastereoisomers.

Step 4:

A mixture of compound P4 (in separate portions of 1.26 g, 0.77 g and1.09 g; 7.15 mmol total) and H₂SO₄ (in separate portions of 24 mL, 15 mLand 19 mL) is allowed to react at 150° C. for 20 minutes. The combinedreaction mixture is allowed to cool slightly and added dropwise toice-water. The mixture is extracted with EtOAc (3×), washed with brine(1×), dried (Na₂SO₄), filtered and concentrated in vacuo. The residue(0.69 g, 1.76 mmol) is dissolved in EtOH (25 mL) and to this solution isadded POCl₃ (2.4 mL; 27 mmol). The reaction is heated at reflux for 1 h,then poured into ice-water and extracted with CH₂Cl₂ (3×). The organicphase is washed with brine, dried (MgSO₄), filtered and concentrated andthe residue is purified by chromatography to afford compound P5.

Step 5:

A solution of (Boc)₂O (1M in THF, 2.47 mL, 2.47 mmol) is added to asolution of H₂NCH₂CH₂Br.HBr (506 mg, 2.47 mmol) and Et₃N (860 μL, 6.175mmol) in THF (10 mL). The reaction mixture is stirred at RT for 18 h andis partitioned between EtOAc (100 mL) and saturated aqueous NaHCO₃ (25mL). The organic phase is washed with brine, dried over anhydrous MgSO₄and concentrated. The residue is purified by chromatography (5% to 20%EtOAc/hexanes) to give BocNHCH₂CH₂Br. To a cooled solution (0° C.) ofcompound P5 (200 mg, 0.495 mmol) in DMF (3 mL) is added KOtBu (67 mg,0.598 mmol). The mixture is stirred for 15 min, then a solution ofBocNHCH₂CH₂Br (160 mg, 0.717 mmol) in DMF (2 mL) is added. The reactionmixture is stirred at RT for 18 h. Water (1 mL) is added, the mixturewas diluted with EtOAc (100 mL) and the organic phase is washed withsaturated aqueous NaHCO₃ (25 mL) and brine, dried over anhydrous MgSO₄and concentrated. The residue is purified by chromatography (10% to 30%EtOAc/hexanes) to give compound P6.

Step 6:

To a solution of compound P6 (96 mg, 0.175 mmol) in DMSO (2.5 mL) isadded 5N NaOH (175 μL, 0.875 mmol). The mixture is stirred for 30 minand purified by semi preparative HPLC to afford the Boc-deprotectedcarboxylic acid. The compound is treated with Boc₂O in the presence ofNaOH to provide compound P7 as a racemic mixture. Separation by chiralHPLC, using a ChiralCel OD-R column (20×250 mm from Chiral TechnologiesInc) and an isocratic solvent system of 20% H₂O (containing 0.06% TFA)and 80% of a solvent mixture composed of 75% MeCN in H₂O (containing0.06% TFA) provides the (S)-enantiomer P7.

Step 7:

To a mixture of compound P7 (9.2 mg, 0.017 mmol) and CH₂Cl₂ (1.5 mL) isadded TFA (750 μL). The mixture is stirred at RT for 1 h andconcentrated. The residue is dissolved in CH₂Cl₂ (1.0 mL) and to thismixture is added Et₃N (7 μL, 0.051 mmol), followed by EZ-Link™TFP-PEO-biotin (Pierce; 17.2 mg, 0.025 mmol). The reaction mixture isstirred at RT for 18 h, the solvent is evaporated and the residue ispurified by semi-preparative HPLC to afford biotinylated Probe I.

Probe I is determined to have a dissociation constant (Kd) of 1 μM, asmeasured by Isothermal Titration Calorimetry (ITC) according to knownmethods, for example as described in Shaw-Reid et al. J Biol. Chem.278(5):2777-80 (2003).

His-tamed HIV integrase: His-tagged integrase is cloned and expressed ina similar manner as outlined in Barsov et al., J. Virol., Vol 70, No. 7,4484-4494, (1996). Briefly, the integrase gene is PCR amplified from aplasmid containing the HXB2 provirus using forward and reverse primersthat span the first and final codons of integrase, respectively. Primerscontain a 5′ NdeI (forward primer) and 3′ XhoI site (reverse primer)which allows for cloning of the NdeI/XhoI fragment into the Pet28abacterial expression vector (Novagen). DH5a E. coli cells are used togenerate and propagate the DNA vectors while BL21 pLysS E. coli cellsare used for expression of the His-tagged protein.

His-tagged HIV-1 integrase is expressed in BL21 pLysS cells (Stratagene)to an O.D. of 1.4 in a 30 L fermentor at 37° C. and induced with 0.5 mMIPTG for 3 hours at 37° C. The bacterial pellet is re-suspended infreshly prepared extraction buffer (20 mM NaPi pH 5.8, 1 M NaCl, 1 Murea, 1 mM TCEP, 20 mM imidazole, 1 mM PMSF, and 1.5 mL Sigma proteaseinhibitor cocktail for 35 mg pellet) and sonicated. The cell lysate issubjected to centrifugation at 100,000 G for 30 min and the supernatantis loaded onto a HiTrap Ni²⁺ column. The column is washed with theextraction buffer containing 100 mM imidazole and eluted in a 15 minlinear gradient to 1 M imidazole. The His-Integrase fractions are pooledand diluted 1 to 8 in fresh S column buffer (20 mM Na Pi pH 5.8, 1 mMTCEP, 1 mM EDTA, 1 M urea, 10% glycerol) and loaded onto a HiTrap Scolumn. The column is washed with S column buffer then eluted in an 80min linear gradient from 0 to 1 M NaCl. Protein fractions are run on anSDS-PAGE gel and the most concentrated samples without contaminant bandsare pooled and precipitated with 60% ammonium sulfate. The precipitateis then re-suspended in storage buffer (20 mM Hepes pH 7.5, 500 mM NaCl,10% glycerol, 0.5 mM TCEP) then loaded onto a SD200 gel filtrationcolumn. The His-tagged Integrase containing fractions are pooled, dosedfor protein concentration and stored at −80° C. in aliquots. TheHis-tagged integrase samples are thawed on ice and diluted to thedesired concentration in assay buffer for use in the integrasedisplacement assay.

Homogenous Time Resolved Fluorescence (HTRF) Assay System

The relative binding affinity of compounds of the invention to the HIVintegrase target is assessed using a HTRF system based on fluorescenceresonance energy transfer (FRET) between europium cryptate (EuK) asenergy donor and cross linked allophycocyanin (XL665) as acceptor. EuKlabeled streptavidin (CysBio) binding to Probe I and XL665 labeledanti-His antibody (CysBio) binding to His-integrase are used in thesystem. Interaction between integrase and Probe I is monitored by energytransfer between the EuK and XL665 in the integrase-probe complex.Displacement of the biotinylated probe from integrase by a compound ofthe invention results in a loss of fluorescence from the XL665 labeledantibody.

Assay solutions are prepared using an assay buffer consisting of 50 mMHEPES (pH 7.5); 50 mM NaCl; 150 mM KF; 1 mg/ml BSA, 1.0 mM TCEP, 0.05%Tween 20. Assay solution A is prepared with Probe I (30 nM) andHis-integrase (300 nM) in assay buffer with 4.5% DMSO. Assay solutions Bis prepared with a compound of the invention diluted to 120 μM in assaybuffer containing 4.5% DMSO, then serial diluted 11 times in 2-foldsteps of the same buffers. Finally, assay solution C is prepared withStreptavidin-EuK and Anti-his XL665 premixed at concentrations of 6 nMand 150 nM respectively in assay buffer.

The reaction mixture was prepared by adding 5 μL of each assay solution(solutions A and C with one of the serial dilutions of compound insolution B for each reaction mixture) to a 384-well black round bottom,low volume NBS plates (Corning catalog #3676) giving a finalconcentration of 100 nM His-integrase, 10 nM probe, compound ofinvention (40 μM down to 37.5 nM), 50 nM anti-his XL665, 2 nM Strep-EuKand 3% DMSO with a total volume of 15 μL. The reaction mixture wasincubated at room temperature for one hour. The fluorescence was read ona Victor 1420 Multilable HTS reader at 615 nm and 665 nm.

The compounds of the invention have greater affinity for the HIVintegrase target than that of Probe I. However, the increased avidity ofthe biotinylated probe due to tetramerization with Strep-EuK allows theevaluation of compounds of the invention that have significantlystronger association with HIV integrase.

Example 57 C8166 HIV-1 Luciferase Assay (EC₅₀)

C8166 cells are derived from a human T-lymphotrophic virus type 1immortalized but nonexpressing line of cord blood lymphocytes (NIH AIDSreagent 404) and are highly permissive to HIV-1 infection. The pGL3Basic LTR/TAR plasmid is made by introducing the HIV-1 HxB2 LTR sequencefrom nucleotide −138 to +80 (Sca1-HindIII) upstream of the luciferasegene in the pGL3 Basic Vector (a promoterless luciferase expressionvector from Promega catalogue #E1751) with the gene for blasticidineresistance cloned in. The reporter cells are made by electroporatingC8166 cells with pGL3 Basic LTR/TAR and selecting positive clones withblasticidine. Clone C8166-LTRluc #A8-F5-G7 was selected by 3 consecutiverounds of limiting dilution under blasticidine selection. Cultures aremaintained in complete media (consisting of: Roswell Park MemorialInstitute medium (RPMI) 1640+10% FBS+10⁻⁶ M β-mercaptoethanol+10 μg/mLgentamycin) with 5 μg/mL blasticidine, however, blasticidine selectionis removed from the cells before performing the viral replication assay.

Luciferase Assay Protocol Preparation of Compounds

Serial dilutions of HIV-1 inhibitor compounds are prepared in completemedia from 10 mM DMSO stock solutions. Eleven serial dilutions of 2.5×are made at 8× desired final concentration in a 1 ml deep well titerplate (96 wells). The 12^(th) well contains complete media with noinhibitor and serves as the positive control. All samples contain thesame concentration of DMSO 0.1% DMSO). A 25 μL aliquot of inhibitor isadded, to triplicate wells, of a 96 well tissue culture treated clearview black microtiter plate (Corning Costar catalogue #3904). The totalvolume per well is 200 μL of media containing cells and inhibitor. Thelast row is reserved for uninfected C8166 LTRluc cells to serve as thebackground blank control and the first row is media alone.

Infection of Cells

C8166 LTRluc cells are counted and placed in a minimal volume ofcomplete RPMI 1640 in a tissue culture flask (ex. 30×10⁶ cells in 10 mLmedia/25 cm² flask). Cells are infected with HIV-1 or virus with variantintegrase generated as described below at a molecules of infection (moi)of 0.005. Cells are incubated for 1.5 h at 37° C. on a rotating rack ina 5% CO₂ incubator and re-suspended in complete RPMI to give a finalconcentration of 25,000-cells/175 μL. 175 μL of cell mix is added towells of a 96 well microtiter plate containing 25 μL 8× inhibitors.25,000 uninfected C8166-LTRluc cells/well in 200 μL complete RPMI areadded to the last row for background control. Cells are incubated at 37°C. in 5% CO₂ incubator for 3 days.

Luciferase Assay

50 μL Steady Glo (luciferase substrate T_(1/2)=5 hours Promega catalogue#E2520) is added to each well of the 96 well plate. The relative lightunits (RLU) of luciferase is determined using the LUMIstar Galaxyluminometer (BMG LabTechnologies). Plates are read from the bottom for 2seconds per well with a gain of 240.

The level of inhibition (% inhibition) of each well containing inhibitoris calculated as follows:

${\% \cdot \mspace{14mu} {inhibition}} = {\left( {1 - \left\lbrack \frac{{{RLU} \cdot {well}} - {{RLU} \cdot {blank}}}{{{RLU} \cdot {control}} - {{RLU} \cdot {blank}}} \right\rbrack} \right)*100}$

The calculated % inhibition values are used to determine EC₅₀, slopefactor (n) and maximum inhibition (I_(max)) by the non-linear regressionroutine NLIN procedure of SAS using the following equation:

${\% \cdot {inhibition}} = \frac{I_{\max} \times \lbrack{inhibitor}\rbrack^{n}}{\lbrack{inhibitor}\rbrack^{n} + {IC}_{50}^{n}}$

Compounds of the invention tested in the cellular assay described aboveare particularly effective at inhibiting HIV integrase. Compounds ofTable 1 were found to have EC₅₀ values of 300 nM or less. Furthermore,compounds of the invention show unexpected potency across major HIV-1virus HIV variants. Compounds of Table 1 show unexpected potency in atleast four or in all six of the known variants at residues 124 and 125of HIV-1 virus from infected patients, namely Thr124/Thr125,Ala124/Thr125, Ala124/Ala125, Thr124/Ala125, Asn124/Thr125 andAsn124/Ala125. The results of representative compounds are shown inTable 2.

TABLE 2 EC₅₀ (nM) EC₅₀ (nM) EC₅₀ (nM) EC₅₀ (nM) Compound A124/T125T124/T125 T124/A125 A124/A125 1008 11 59 49 55 1010 12 41 76 49 1014 1987 76 68 1018 35 55 93 110 1023 12 29 31 37 1038 3 9 15 8 1052 28 120 5964 1059 3 14 11 9 1136 91 200 140 170 2001 71 110 120 67 2002 9.9 19 2413Generation of Viruses with 124/125 Variant Residues of Integrase

The 2.12 virus with NL4.3 strain of HIV-1 integrase (SEQ ID NO: 1) isused for the Thr124/Thr125 variant integrase residues and HXB2 integraseis introduced into the 2.12 virus for the Ala124/Thr125 variantintegrase residues. The remaining variants viruses are generated by sitedirected mutagenesis of NL4.3 integrase to introduce the Ala124/Ala125,Thr124/Ala125, Asn124/Thr125, or Asn124/Ala125 variants in the 2.12virus. Molecular biology and generation of virus are performed in asimilar manner to Doyon et al. J. Virol. 70(6):3763-9 (1996). Briefly,site directed mutagenesis is used to introduce silent mutations in theintegrase gene of the 2.12 virus. The mutations in NL4.3 integraseintroduced unique Cla I and Xba I restriction sites in the 2.12 provirus(using the primers 5′-TTT AGA TGG AAT CGA TAA GGC CCA AGA AG-3′ and5′-CTT CTT GGG CCT TAT CGA TTC CAT CTA AA-3′ to introduce Cla I site and5′-GGT TTA TTA CAG GGA CTC TAG AGA TCC AGT TTG GA-3′ and 5′-TCC AAA CTGGAT CTC TAG AGT CCC TGT AAT AAA CC-3′ to introduce XbaI site). The 2.12virus is generated with the Cla I and Xba I restriction sites inintegrase replicates and responds to compounds of the invention in asimilar manner to the original 2.12 virus. Point mutations at residues124 and 125 are introduced into NL4.3 integrase in a Pet 28a vector(Novagen) by using the QuikChange site-directed mutagenesis kit(Stratagene). After sequencing to ensure that the desired 124 and/or 125residue mutations are obtained, the Cla I/Xba I fragment is PCRamplification with Pfu Ultra (Stratagene) and cloned into the unique ClaI/Xba I sites in the 2.12 virus.

Tables of Compounds

The following tables list compounds of the invention. Compounds of theinvention tested in the cellular assay described above in Example 57 areparticularly effective at inhibiting HIV integrase. Compounds of Table 1were found to have EC₅₀ values of 300 nM or less in at least four or inall six of the known variants at residues 124 and 125 of HIV-1 virusfrom infected patients, namely Thr124/Thr125, Ala124/Thr125,Ala124/Ala125, Thr124/Ala125, Asn124/Thr125 and Asn124/Ala125.

Retention times (t_(R)) for each compound are measured using thestandard analytical HPLC conditions described in the Examples. As iswell known to one skilled in the art, retention time values aresensitive to the specific measurement conditions. Therefore, even ifidentical conditions of solvent, flow rate, linear gradient, and thelike are used, the retention time values may vary when measured, forexample, on different HPLC instruments. Even when measured on the sameinstrument, the values may vary when measured, for example, usingdifferent individual HPLC columns, or, when measured on the sameinstrument and the same individual column, the values may vary, forexample, between individual measurements taken on different occasions.

TABLE 1

 Cpd R⁴ R⁶ R⁷ t_(R) (min) MS (M + H)⁺ 1001

CH₃ H 4.7 398.1/400.1 1002

H CH₃ 4.6 398.1/400.1 1003

H F 4.5 402.2/404.1 1004

H H 3.9 396.2 1005

H H 5.1 404.2 1006

H H 4.3 406.2 1007

H H 4.5 364.2 1008

H H 4.8 378.2 1009

H H 4.6 382.1 1010

H H 4.8 402.1/404.1 1011

H H 4.7 406.2 1012

H H 4.7 428.0/430.0 1013

H H 3.9 442.1 1014

H H 3.7 392.1 1015

H H 5.0 398.1/400.1 1016

H H 5.6 418.2 1017

H H 5.0 420.2 1018

H H 3.7 400.1 1019

H H 3.7 382.1 1020

H CH₃ 4.0 413.2 1021

H CH₃ 4.3 420.1 1022

F H 4.9 424.2 1023

H H 4.4 390.1 1024

H H 5.2 420.1/422.1 1025

H CH₃ 4.4 364.2 1026

H CH₃ 5.3 432.1/434.1/436.1 1027

H CH₃ 5.5 406.2 1028

H CH₃ 3.6 415.2 1029

H CH₃ 5.1 460.1/462.1 1030

H CH₃ 4.4 416.1/418.2 1031

H CH₃ 4.8 396.2 1032

H CH₃ 4.8 430.2 1033

H CH₃ 5.3 454.1/456.1 1034

H CH₃ 4.6 404.2 1035

H H 4.9 398.1/400.1 1036

H CH₃ 4.8 410.2 1037

H H 5.3 454.2/456.2 1038

H H 4.9 440.2/442.2 1039

H H 5.1 440.2/442.2 1040

H H 4.1 424.1 1041

H CH₃ 4.3 438.2 1042

H CH₃ 5.6 468.1/470.1 1043

CH₃ H 5.2 434.2 1044

H H 4.9 420.2 1045

CH₃ H 4.4 416.1/418.1 1046

H H 4.8 398.1/400.1 1047

H H 4.7 424.1 1048

H H 4.5 424.1/426.1 1049

H H 4.8 424.1/426.1 1050

H H 3.9 390.1 1051

H CH₃ 4.3 416.1/418.1 1052

H H 4.1 420.2 1053

CH₃ H 5.1 454.1/456.1 1054

CH₃ H 5.4 454.1/456.1 1055

H CH₃ 5.2 454.1/456.1 1056

H CH₃ 5.4 454.1/456.1 1057

CH₃ H 5.1 438.3 1058

CH₂CH₃ H 5.5 412.2/414.2 1059

H H 4.9 438.2 1060

H H 3.7 424.4 1061

H H 3.9 442.2 1062

H H 4.5 426.2/428.2 1063

H H 3.7 406.2 1064

H H 4.8 436.3 1065

H H 3.8 440.2/442.1 1066

H H 4.6 406.2 1067

H H 4.1 440.2 1068

H H 4.9 420.2 1069

H H 4.6 406.2 1070

H H 4.4 434.1/436.1 1071

H H 5.0 424.1/426.1 1072

H H 4.7 424.1/426.1 1073

H H 4.9 424.1/426.1 1074

H H 5.0 396.2 1075

H H 3.6 415.3 1076

H CH₃ 3.9 429.2 1077

H H 3.8 421.2 1078

H CH₃ 5.7 448.1/450.1 1079

H H 5.2 442.2 1080

H H 5.4 440.1 1081

H H 4.6 398.2 1082

H CH₃ 4.9 403.2 1083

F H 5.6 458.1/460.1 1084

H CH₃ 4.5 449.2/451.2 1085

H CH₃ 3.4 429.3 1086

H H 5.0 426.2/428.2 1087

H H 4.8 428.2/430.2 1088

H H 4.8 424.1/426.1 1089

H H 4.9 416.2/418.2 1090

F H 5.5 438.2 1091

Cl H 4.8 455.2/457.2 1092

Cl H 5.4 454.2/456.2 1093

H H 4.1 434.2 1094

H H 5.5 420.2 1095

H CH₃ 4.6 435.2 1096

CH₃ CH₃ 5.3 448.3 1097

H H 4.3 455.2/457.2 1098

H H 5.5 476.1/478.1 1099

F H 4.4 439.2 1100

CH₃ H 4.5 435.2 1101

H H 3.9 432.2 1102

H H 4.4 441.2/443.1 1103

H H 4.1 419.3 1104

H H 4.5 440.2/442.1 1105

H H 5.1 458.2/460.2 1106

Cl H 5.8 492.1/494.1/496.1 1107

Cl H 5.6 474.2/476.2 1108

F H 4.2 459.2/461.1 1109

Cl H 4.7 475.1/477.1/479.1 1110

CH₃ H 4.6 455.2/457.2 1111

H CH₃ 4.5 455.2/457.2 1112

H H 5.0 436.1/438.1/440.1 1113

H H 5.4 432.1/434.1/438.1 1114

H H 4.4 384.1/386.1 1115

H H 4.3 449.3 1116

H H 4.4 469.2/471.2 1117

H H 4.5 402.1/404.1 1118

F CH₃ 4.4 453.2 1119

F CH₃ 5.4 473.2/475.2 1120

H H 4.6 402.1/404.1 1121

H H 4.4 382.2 1122

F CH₃ 5.5 452.3 1123

H CH₃ 5.3 494.0/496.0/498.0 1124

H H 4.5 402.2/404.2 1125

H H 5.1 480.1/482.1/484.1 1126

Cl H 5.9 514.0/516.0/518.0 1127

H H 4.9 416.2/418.2 1128

F H 5.4 498.0/500.0/502.0 1129

H H 5.3 442.2/444.2 1130

H H 4.9 460.1/462.1 1131

H —CH₃ 3.6 457.3 1132

H CH₃ 5.2 450.1/452.1/454.1 1133

F H 5.4 454.1/456.1/458.1 1134

F H 5.2 434.2/436.2 1135

Cl H 5.6 450.1/452.1/454.1 1136

H H 3.0 407.1 1137

H Me 3.7 471.3 1138

H Me 5.0 463.2/465.2 1139

H Me 4.8 468.2/469.2 1140

H Me 4.4 447.3 1141

H Me 3.1 441.2 1142

H Cl 5.4 475.1/477.1/479.1 1143

H Cl 3.1 477.2/479.2 1144

H H 3.7 443.2 1145

H H 3.2 441.3 1146

H H 4.1 433.3 1147

H H 3.8 457.2 1148

H H 2.8 472.2 1149

H H 4.5 430.0 1150

Me H 3.7 457.2 1151

Cl H 3.0 477.3/479.3 1152

F H 2.8 461.3 1153

F Me 2.9 475.1

TABLE 2

t_(R) MS Cpd R³ R⁴ (min) (M + H)⁺ 2001

3.0 429.2 2002

2.9 457.3

Each of the references including all patents, patent applications andpublications cited in the present application is incorporated herein byreference in its entirety, as if each of them is individuallyincorporated. Further, it would be appreciated that, in the aboveteaching of invention, the skilled in the art could make certain changesor modifications to the invention, and these equivalents would still bewithin the scope of the invention defined by the appended claims of theapplication.

1-38. (canceled)
 39. A compound of the formula (I)

wherein R⁴ is aryl or Het, wherein each of the aryl and Het isoptionally substituted with 1 to 3 substituents each independentlyselected from halo, (C₁₋₆)alkyl, (C₂₋₆)alkenyl, (C₁₋₆)haloalkyl,(C₃₋₇)cycloalkyl, —OH, —O(C₁₋₆)alkyl, —SH, —S(C₁₋₆)alkyl, —NH₂,—NH(C₁₋₆)alkyl and —N((C₁₋₆)alkyl)₂; wherein the (C₁₋₆)alkyl isoptionally substituted with hydroxy, cyano or oxo; R⁶ and R⁷ are eachindependently selected from H, halo, (C₁₋₆)alkyl and (C₁₋₆)haloalkyl;wherein Het is a 4- to 7-membered saturated, unsaturated or aromaticheterocycle having 1 to 4 heteroatoms each independently selected fromO, N and S, or a 7- to 14-membered saturated, unsaturated or aromaticheteropolycycle having wherever possible 1 to 5 heteroatoms, eachindependently selected from O, N and S; or a salt thereof.
 40. Acompound according to claim 39, or a pharmaceutically acceptable saltthereof, wherein R⁴ is Het optionally substituted with 1 to 2substituents each independently selected from halo, (C₁₋₃)alkyl andO—(C₁₋₃)alkyl.
 41. A compound according to claim 40, or apharmaceutically acceptable salt thereof, wherein R⁴ is Het optionallysubstituted with 1 to 2 substituents each independently selected fromCl, F, CH₃ and CH₂CH₃ wherein said Het is defined as a 7- to 14-memberedsaturated, unsaturated or aromatic heteropolycycle having whereverpossible 1 to 2 heteroatoms, each independently selected from O, N andS.
 42. A compound according to claim 39, or a pharmaceuticallyacceptable salt thereof, wherein R⁴ is selected from:

optionally substituted 1 to 3 times with halo, (C₁₋₃)alkyl andO—(C₁₋₃)alkyl.
 43. A compound according to claim 42, or apharmaceutically acceptable salt thereof, wherein R⁴ is selected from:

optionally substituted 1 to 2 times with halo, (C₁₋₃)alkyl andO—(C₁₋₃)alkyl.
 44. A compound according to claim 39, or apharmaceutically acceptable salt thereof, wherein R⁶ is H, F, Cl or(C₁₋₂)alkyl.
 45. A compound according to claim 44, or a pharmaceuticallyacceptable salt thereof, wherein R⁶ is H or CH₃.
 46. A compoundaccording to claim 39, or a pharmaceutically acceptable salt thereof,wherein R⁷ is H, F, Cl or CH₃.
 47. A compound according to claim 46, ora pharmaceutically acceptable salt thereof, wherein R⁷ is H or CH₃. 48.A compound according to claim 39 having the following formula:

wherein R⁴, R⁶ and R⁷ are defined as: R⁴ R⁶ R⁷

CH3 H;

H CH3;

H F;

H H;

H H;

H H;

H H;

H H;

H H;

H H;

H H;

H H;

H H;

H H;

H H;

H H;

H H;

H H;

H H;

H CH₃;

H CH₃;

F H;

H H;

H H;

H CH₃;

H CH₃;

H CH₃;

H CH₃;

H CH₃;

H CH₃;

H CH₃;

H CH₃;

H CH₃;

H CH₃;

H H;

H CH_(3;)

H H;

H H;

H H;

H H;

H CH₃;

H CH₃;

CH₃ H;

H H;

CH₃ H;

H H;

H H;

H H;

H H;

H H;

H CH₃;

H H;

CH₃ H;

CH₃ H;

H CH₃;

H CH₃;

CH₃ H;

CH₂CH₃ H;

H H;

H H;

H H;

H H;

H H;

H H;

H H;

H H;

H H;

H H;

H H;

H H;

H H;

H H;

H H;

H H;

H H;

H CH₃;

H H;

H CH₃;

H H;

H H;

H H;

H CH₃;

F H;

H CH₃;

H CH₃;

H H;

H H;

H H;

H H;

F H;

Cl H;

Cl H;

H H;

H H;

H CH₃;

CH₃ CH₃;

H H;

H H;

F H;

CH₃ H;

H H;

H H;

H H;

H H;

H H;

Cl H;

Cl H;

F H;

Cl H;

CH₃ H;

H CH₃;

H H;

H H;

H H;

H H;

H H;

H H;

F CH₃;

F CH₃;

H H;

H H;

F CH₃;

H CH₃;

H H;

H H;

Cl H;

H H;

F H;

H H;

H H;

H —CH₃;

H CH₃;

F H;

F H;

Cl H; or

H H;

or a pharmaceutically acceptable salt thereof.
 49. A compound accordingto claim 39 having the following formula:

wherein R⁴, R⁶ and R⁷ are defined as: R⁴ R⁶ R⁷

H CH₃;

H CH₃;

H CH₃;

H CH₃;

H CH₃;

H Cl;

H Cl;

H H;

H H;

H H;

H H;

H H;

H H;

CH₃ H;

Cl H;

F H; or

F CH₃

or a pharmaceutically acceptable salt thereof.
 50. A pharmaceuticalcomposition comprising a compound of formula (I) according to any one ofclaims 39 to 49 or a pharmaceutically acceptable salt thereof; and apharmaceutically acceptable carrier.
 51. A method for treating HIVinfection in a host infected by HIV which method comprises administeringto such host a therapeutically effective amount of a compound of formula(I) according to any one of claims 39 to 49 or a pharmaceuticallyacceptable salt thereof.